TW202115077A - Malt1 inhibitors and uses thereof - Google Patents

Abstract

The invention provides a compound represented by structural formula (I): or a pharmaceutically acceptable salt thereof useful for treating an autoimmune disorder, an inflammatory disease, or a cancer.

Description

MALT1 inhibitor and its use

Cross-reference to related applications

This application claims the benefit of the priority of the International Patent Application No. PCT/CN2019/094154 filed on July 1, 2019. The entire content of the aforementioned application is incorporated herein by reference in its entirety.

The present invention relates to the use of novel compounds or pharmaceutically acceptable salts thereof to treat autoimmune diseases, inflammatory diseases, or cancer.

Paracaspase is a member of the C14 family of cysteine proteases. Compared with metacaspases that exist in plants, fungi, and protists, they are related to animals and slime molds. Protease related to apoptosis protease. The para-apoptotic protease is more similar to the apoptotic protease than the meta-apoptotic protease, indicating that the para-apoptotic protease system deviates from the common meta-apoptotic protease ancestor.

Para-apoptotic protease was first identified in the recurrent t(11;18)(q21;q21) chromosomal translocation associated with a subset of MALT (gastric mucosa-associated lymphoid tissue) lymphoma. This chromosomal translocation is the most common mutation identified in MALT lymphoma, and the translocation results from the apoptosis protease inhibitor protein c-IAP2 (also It is called the baculovirus IAP repeat sequence protein 3) The formation of a fusion oncoprotein composed of the N-terminus and the C-terminus of the human para-apoptotic protease is called MALT1.

MALT1 (Mucosa Associated Lymphoid Tissue lymphoma translocation protein 1, gastric mucosa-associated lymphoma lymphoma translocation protein 1) is the only para-apoptotic protease in humans. MALT1 has both scaffold and protease functions. It also transmits messages from cell surface receptors in both immune cells (such as B and T lymphocytes, NK lymphocytes, and myeloid cells and mast cells) and non-immune cells. . In immune cells, MALT1 uses ITAM (Immunoreceptor Tyrosine-based Activation Motif, immune receptor tyrosine activation motif) sequence in immune receptors [for example, cell receptor (BCR) and T cell receptor (TCR)] Play downstream. In non-immune cells, MALT1 acts downstream of specific G protein-coupled receptors (GPCR) and EGFR (Her2/neu). The scaffold function of MALT1 is to help other communication proteins assemble into a communication complex called CBM (see below), and its cysteine protease function, at least in some immune cells, is responsible for cleaving some targeted proteins. The proteolytic activity of MALT1 seems to be required for T cell activation and the development of B cell lymphoma.

For example, after TCR/BCR activation, MALT1 becomes the active subunit of the so-called CBM complex, which consists of multiple subunits of three proteins: CARD11 (capspace recruitment domain family member 11, apoptotic protease aggregation domain Family member 11, also known as CARMA1), BCL10 (B-cell CLL/lymphoma 10) and MALT1. However, depending on the cell lineage, MALT1 and its partner BCL10 usually bind to different members of CARMA (CARD-containing membrane-associated guanylate kinase) protein family containing CARD (caspace recruitment domain, apoptotic protease aggregation domain). In particular, in lymphocytes, the CBM complex formed immediately after antigen receptor stimulation (via the TCR or BCR pathway) involves CARMA1/CARD11, and CARD9 interacts with MALT1 downstream of Toll-like or C-type lectin receptors. Meanwhile, in non-immune cells, the CBM complex MALT1- BCL10-CARD10 via the GPCR and the activation of NF- κ B connected messaging (McAllister-Lucas et al, PNAS 104:. 139-44,2007). On the other hand, CARD14 interacts with MALT1 and BCL10 in keratinocytes.

MALT1 has an N-terminal death domain (DD) with unknown function, followed by two immunoglobulin-like domains (Ig) required for BCL10 binding. The central apoptotic protease domain shares low sequence homology with the apoptotic protease, but adopts a folded structure that is significantly similar to that of the apoptotic protease, with the active site residues of H415 and C464. The apoptotic protease domain is followed by the third Ig domain containing K644, which is the site of monoubiquitination that controls protease activity. MALT1 also contains two ubiquitin ligase TRAF6 (tumor necrosis factor receptor-associated factor 6, tumor necrosis factor receptor-associated factor 6) binding motifs. TRAF6 polyubiquitinates MALT1 on multiple C-terminal lysine residues to produce a K63-linked ubiquitin chain, which can then promote NF through the recruitment of IKK-activated kinase TAK1 through the aggregation of the adaptor protein TAB 2/3 -Activation of κB kinase (IKK) complex inhibitors.

MALT1 is a key mediator of the typical NF-κB message transmission pathway, and acts as a core protein in many diseases that directly or indirectly involve the inflammatory transcription factor NF-κB. MALT1 affects NF-κB signaling through two different mechanisms. First, through its scaffolding function, MALT1 aggregates NF-κB signaling proteins such as TRAF6, TAB-TAK1, or NEMO-IKK α / β . Second, through its Cys protease function, MALT1 cleaves and inactivates the negative regulators of NF-κB signaling such as RelB, A20, or CYLD. The ultimate indicators of MALT1 activity are the nuclear translocation of the NF-κB transcription factor complex and the activation of NF-κB signaling (Jaworski et al. , Cell Mol Life Science 73:459-473, 2016), leading to the production of interleukin-2 (IL-2), and activation and proliferation of T and B lymphocytes.

The c-IAP2-MALT1 fusion is a powerful activator of the NF-κB pathway (Rosebeck et al. , World J Biol Chem 7: 128-137, 2016). The c-IAP2-MALT1 fusion mimics the ligand-binding TNF receptor and promotes TRAF2-dependent ubiquitination of RIP1 that acts as a scaffold for activating canonical NF-κB signaling. Furthermore, it has been shown that the cIAP2-MALT1 fusion can cleave and produce stable and constitutively active fragments of NF-κB-inducible kinase (NIK), thereby activating the atypical NF-κB pathway (Rosebeck et al. , Science 331: 468-472, 2011).

MALT1 functions as an intracellular signaling protein in congenital [eg, natural killer or NK cells, dendritic cells (DC), and mast cells] and adaptive immune cells (eg, T cells and B cells) . Some publications seem to suggest that MALT1 and its proteolytic function are in many cell types in the signaling cascade triggered by innate cell receptors such as Dectin receptors, and the signaling cascade triggered by G protein-coupled receptors (GPCRs) An important role in the reaction. Therefore, MALT1 has attracted much attention in the mechanism of autoimmunity and inflammatory pathology. In addition, it is noted that persistent (dysregulated) MALT1 activity is associated with MALT lymphoma and activated B cell-like diffuse large B cell lymphoma (ABC-DLBCL).

Indeed, the continuous activation of NF-κB messaging is the quality assurance of ABC-DLBCL. DLBCL (diffuse large B-cell lymphoma) is a B-cell cancer, the most common form of adult non-Hodgkin’s lymphoma (NHL), accounting for about 25% of lymphoma cases. DLBCL is an aggressive tumor that can appear in almost any part of the body. Normally, DLBCL is caused by normal B cells, but it can also represent other types of lymphomas or malignant transformations of leukemias with potentially significant risk factors for immunodeficiency. In biology, there are two different molecular subtypes of DLBCL: germinal center B cells (GCB) and activated B cells (ABC). ABC-DLBCL is derived from B cells during the differentiation of germinal center B cells into plasma cells. Approximately 40% of all DLBCL is ABC-DLBCL, which is a more radical form of DLBCL. Generally, patients diagnosed with ABC subtype have a worse prognosis than GCB patients. The activation of the NF-κB pathway in ABC-DLBCL is driven by mutations in signaling components (such as CD79A/B, CARD11, MYD88, or A20) (Staudt, Cold Spring Harb Perspect Biol 2010, 2; Lim et al. , Immunol Rev 246: 359-378, 2012). Despite advances in treatment, one-third of DLBCL patients still fail to respond or relapse within a short period of time.

MALT1 has also been reported to be involved in several other disease pathologies. For example, different types of tumor diseases include lung adenocarcinoma (Jiang et al. , Cancer Research 71: 2183-2192, 2011; Pan et al. , Oncogene 1: 10, 2015), breast cancer (Pan et al. , Mol Cancer Res 14: 93-102, 2016), adventitia cell lymphoma (Penas et al. , Blood 115: 2214-2219, 2010; Rahal et al. , Nature Medicine 20 : 87-95, 2014), marginal band lymphoma (Remstein et al. , Am J Pathol 156: 1183-1188, 2000; Baens et al. , Cancer Res 66: 5270-5277, 2006; Ganapathi et al. , Oncotarget 1:10,2016; Bennett et al. , Am J of Surgical Pathology 1:7,2016), cutaneous T-cell lymphoma [e.g. Sezary syndrome] (Qin et al. , Blood 98: 2778-2783 ,2001; Doebbeling et al. , J of Exp and Clin Cancer Res 29:1-5,2010), primary exudate lymphoma (Bonsignore et al. , Leukemia 31:614-624,2017), pancreatic cancer (WO2016/193339A1), a specific type of chronic lymphocytic leukemia with CARD11 mutation, and a specific subtype of the GCB-DLBCL type of lymphoma involving MALT1.

In addition to lymphoma, MALT1 has been shown to play a key role in innate and adaptive immunity (Jaworski et al. , Cell Mol Life Sci. 2016).

MALT1 scaffold and protease functions are necessary for the development of peritoneal B1 B cells, border zone (MZ) B cells, and natural regulatory T cells (nTreg). The polarization of naive CD4 ⁺ T cells into the Th17 subset of T helper cells is also heavily dependent on the MALT1 protease function.

For example, MALT1 has been found to play a key role in experimental allergic encephalomyelitis (a mouse model of multiple sclerosis) in mice (McGuire et al. , J. Neuroinflammation 11:124, 2014). It has been shown, the onset and progression of experimental allergic encephalomyelitis in mice diseases MALT1 protease inhibitors may be attenuated (McGuire et al, J.Neuroinflammation 11: . 124,2014). Mice exhibiting a catalytically inactive MALT1 mutant showed loss of border zone B cells and B1 B cells, and a general immune deficiency characterized by reduced T and B cell activation and proliferation. However, these mice also developed spontaneous multi-organ autoimmune inflammation at 9 to 10 weeks of age. Why the knock-in mice whose MALT1 protease is invalid show the destruction of tolerance, but the conventional MALT1 KO mice do not, it is still not well understood. One hypothesis suggests that the imbalanced immune homeostasis of MALT1 protease-invalid gene knock-in mice may be due to the incomplete lack of T and B cells, but the severe lack of immunoregulatory cells (Jaworski et al. , EMBO J. 2014; Gewis et al. , Cell Reports 2014; Bornancin et al. , J. Immunology 2015; Yu et al. , PLOS One 2015). Similarly, human MALT1 deficiency is associated with compound immunodeficiency (McKinnon et al. , J. Allergy Clin. Immunol. 133: 1458-1462, 2014; Jabara et al. , J. Allergy Clin. Immunol. 132: 151 -158, 2013; Punwani et al. , J. Clin. Immunol. 35: 135-146, 2015). In view of the difference between gene mutation and pharmacological inhibition, the phenotypes of knock-in mice whose MALT1 protease is invalid may be different from those of patients treated with MALT1 protease inhibitors. The reduction of immunosuppressive T cells by inhibiting the MALT1 protease may be beneficial to cancer patients due to the potential to enhance anti-tumor immunity.

In any case, the dysregulation of MALT1 activity occurs in, for example, MALT1-dependent inflammatory and/or autoimmune diseases [e.g., rheumatoid arthritis (rheumatoid anthritis, RA), multiple sclerosis (MS), psoriasis, systemic Lupus, Sjogren's syndrome, and Hashimoto's thyroiditis] have a role in the formation of diseases. Targeted immunomodulatory proteins can have direct and indirect benefits in various inflammatory diseases of multiple organs, for example in the treatment of psoriasis (Lowes et al. , Ann Review Immunology 32: 227-255, 2014; Afonina et al. , EMBO Reports 1-14,2016; Howes et al. , Biochem J 1-23,2016), multiple sclerosis (Jabara et al. , J Allergy Clin Immunology 132:151-158,2013; McGuire et al. , J of Neuroinflammation 11: 1-12, 2014), rheumatoid arthritis, Sjogren’s syndrome (Streubel et al. , Clin Cancer Research 10: 476-480, 2004; Sagaert et al. , Modern Pathology 19: 225-232, 2006) , Ulcerative colitis (Liu et al. , Oncotarget 1-14, 2016), MALT lymphoma in different organs (Suzuki et al. , Blood 94: 3270-3271, 1999; Akagi et al. , Oncogene 18: 5785- 5794, 1999) and different types of allergic diseases caused by chronic inflammation.

(mepazine)。此等MALT1抑制劑之一特徵是，該等化合物被推薦用於依賴NF-κB路徑活性失調之自體免疫或炎性路徑、或癌症。MALT1蛋白分解活性之其他抑制劑已被敘述在臨床前淋巴瘤模式中具有活性(Vincendeau et al.,Int.J.Hematoi.Oncol.2：409,2013)。 Inhibitors of MALT1 activity have been identified as potential therapeutic drugs. Rebaud et al. ( Nat Immunol 9(3):272-81,2008) describe the matrix analogue zVRPRfmk equipped with warheads, while Lim et al. ( J Med Chem 58(21):8591-8502,2015) describe small molecules MALT1 inhibitor MI2. Nagel et al. ( Cancer Cell 22(6):825-837,2012) described another small molecule inhibitor mepiidine

(mepazine). One of the characteristics of these MALT1 inhibitors is that the compounds are recommended for autoimmune or inflammatory pathways, or cancers that rely on the dysregulation of the NF-κB pathway activity. Other inhibitors of MALT1 proteolytic activity have been described as active in preclinical lymphoma models (Vincendeau et al. , Int. J. Hematoi. Oncol. 2:409, 2013).

Similarly, Novartis (WO2015/181747) disclosed a small molecule MALT1 inhibitor, and used MALT1 biochemical analysis and the NF-κB reporter gene driven by the ectopic expression of the cIAP2-MALT1 fusion protein specific to MALT lymphoma Analysis and analysis of the reporter gene driven by the IL2 promoter identified these compounds. See also WO2017/081641 (Novartis) for a subset of compounds of the same class.

For example, by using verification by Lu Nepal clinical concepts (Ibrutinib)] and the like of a BTK inhibitor showed inhibition of NF- κ B messaging is effective in treating ABC-DLBCL. MALT1 acts downstream of BTK in the NF-κB signaling pathway. Therefore, MALT1 inhibitors can target ABC-DLBCL patients who are non-responsive or have developed resistance to BTK inhibitors (such as ibrutinib), mainly patients with CARD11 mutations.

The efficacy of MALT1 protease small molecule inhibitors has been demonstrated in the preclinical model of ABC-DLBCL (Fontan et al. , Cancer Cell 22: 812-824, 2012; Nagel et al. , Cancer Cell 22: 825-837, 2012; Fontan et al. , Clin Cancer Res 19: 6662-68, 2013). Furthermore, the covalent catalytic sites and ectopic inhibitors of the MALT1 protease function have been described, indicating that this protease inhibitor can be used in pharmaceutical preparations (Demeyer et al. , Trends Mol Med 22: 135-150, 2016).

Therefore, the industry has a demand for MALT1 inhibitory compounds for the treatment of diseases or disorders involving MALT1 activation (such as cancer and immune and inflammatory disorders that rely on MALT1-NF-kB activation).

The compounds of formula (I), (II-A), or (II-B) that inhibit the activity of MALT1, and the compounds of the examples (herein collectively referred to as "compounds of the present invention"), and pharmaceutically acceptable compounds are described herein salt.

In one aspect, the present invention provides a compound represented by structural formula (I):

或其醫藥上可接受之鹽、或立體異構物，式中

Or its pharmaceutically acceptable salt or stereoisomer, where

或

； A ring is

or

；

Z is O, NR ₆ , or S;

A ₁ and A ₂ are each independently CR ₁ or N;

Each case of R ₁ is hydrogen; halogen; -OH; CN; -COOC _1-6 alkyl; if necessary, C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; phenyl; amine Group; N,N -di-C _1-6 alkylamino group; optionally through halogen, phenyl, or a 5- to 6-membered heterocyclic ring with 1 to 3 heteroatoms selected from N and O (the ring is optionally (C _1-6 alkyl substituted) substituted C _1-6 alkyl; Rh; ORh; a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms selected from N and O, which ring may optionally be amine group, optionally via amine or hydroxyl or by N - substituted bis-substituted -C _1-6 alkylaminocarbonylamino C _1-6 alkyl - -C _1-6 alkyl mono aminocarbonyl or N, N; among them

Rh is a 5- to 6-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O and S, and the ring _{is optionally substituted with C 1-6} alkyl, -OH, or pendant oxy;

Each case of R ₂ is hydrogen; halogen; CN; -COOC _1-6 alkyl; optionally C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; amino; N,N- Di-C _1-6 alkylamino group; optionally via halogen, -OH, phenyl, or a 5- to 6-membered heterocyclic ring with 1 to 2 heteroatoms selected from N and O (the ring optionally via C _{1 -6} alkyl substitution) substituted C _1-6 alkyl; Rh; ORh; a 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, which ring may optionally be through an amino group, optionally _{Requires C 1-6} alkyl substituted by amine or hydroxy, or substituted by N -mono-C _1-6 alkylaminocarbonyl or N,N -di-C _1-6 alkylaminocarbonyl;

Each case of R ₃ is H; deuterium; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; -C(=O)NR _a R _b ; optionally halogen, OH, C _1-6 alkyl, -NH ₂ , -NHC(=0)C _1-6 alkyl, N -di-C _1-6 alkylamino, or N- the mono-substituted -C _1-6 alkyl C _1-6 alkoxy group; an optionally halogen, C _{2-6 alkenyl, -OH, -NH 2, -NHC (} = O) C 1-6 alkyl Group, N -di-C _1-6 alkylamino group, N -mono-C _1-6 alkylamino group, -O-Rg, Rg, phenyl, or C _1-6 alkoxy substituted C _{1 -6} alkyl (wherein the alkoxy group is optionally substituted by halogen, -OH, C _1-6 alkoxy, N,N -di-C _1-6 alkylamino, Rg or phenyl); C _3-6 cycloalkane substituted by halogen, -OH, C _1-6 alkyl, N,N -di-C _1-6 alkylamino or C _1-6 alkoxy-C _{1-6 alkyl} Group; phenyl substituted with halo or C _1-6 alkoxy as required; 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, and the ring may be optionally controlled Amino or -OH-substituted C _1-6 alkyl substitution; Rg; or N,N -di-C _1-6 alkylaminocarbonyl; wherein

Rg is a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, and the ring is optionally controlled by -OH, -NH ₂ , C _1-6 alkyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution;

R _a is independently H or an optionally substituted C _1-6 alkoxy group of C _1-6 alkyl, Rb is independently H, C _1-6 alkyl, -COC _1-6 alkyl, -SO ₂ C _1-6 alkyl, C _3-6 cycloalkyl or a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, the ring is optionally connected to C _1-6 alkyl, C _{1- 6} alkoxy-C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution, or

R _a and R _b together with the nitrogen atom to which it is connected, form a 4- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O, and S, and the ring is optionally controlled by -OH, -NH ₂ , N -Di-C _1-6 alkylamino, N-mono-C _1-6 alkylamino, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _{1 -6} haloalkoxy, O-cyclopropyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkyl-carbonyl substitution;

Each example of R _{c is C 1-6} alkyl or C _3-6 cycloalkyl;

Wherein the R _a, R _b, or R _c or as shown by the R _a, R _b, or the R groups depending on the alkyl group shown _c needs halogen, -OH, C _1-2 alkoxy, or C _3-4 cycloalkyl substitution;

Each case of R ₄ is H, deuterium, halogen, CN, C _1-6 alkyl, or C _1-6 haloalkyl;

Each case of R ₄ 'is H, deuterium, F, or Cl;

Each case of R ₅ is H, deuterium, C _1-6 alkyl, or C _1-6 haloalkyl; and

R ₆ is H; OH; optionally substituted with halogen, OH, C _1-6 alkoxy or C _1-6 alkyl; or an optionally halogen-substituted OH, C _1-6 alkoxy groups or C _3-6 cycloalkyl.

Provided herein is a pharmaceutical composition comprising an effective amount of the compound of the present invention or its medical A pharmaceutically acceptable salt and a pharmaceutically acceptable carrier.

Also provided herein is a combination therapy comprising a therapeutically effective amount of a compound of the present invention, or a pharmaceutically acceptable salt thereof, and one or more co-agents with therapeutic activity.

The present invention further provides a method for inhibiting MALT1 activity in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention or a pharmaceutically acceptable salt thereof.

In a specific embodiment of the method of the present invention, the subject has a disease or symptom such as an autoimmune disease, inflammatory disease, or cancer, and wherein the disease or symptom is treated.

In a specific example of the method of the present invention, the disease or symptom is rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE), vasculitis, allergic disease, Respiratory diseases [such as asthma and chronic obstructive pulmonary disease (COPD)], symptoms caused by delayed or immediate allergic reactions, acute allergies, acute or chronic transplant rejection, graft-versus-host disease, cancer of hematopoietic origin, or solid tumors , Including chronic myelogenous leukemia (CML), myelogenous leukemia, non-Hodgkin’s lymphoma (NHL), or B-cell lymphoma.

Specific specific examples disclose the compound of the present invention, or a pharmaceutically acceptable salt thereof, used as a medicament, for example, a medicament of a MALT1 inhibitor.

The present disclosure also provides the use of the compound of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound in any of the above-mentioned methods of the present invention. In a specific example, a compound of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound used in any of the above methods of the present invention is provided. In another specific example, the use of the compound of the present invention or a pharmaceutically acceptable salt thereof or a pharmaceutical composition containing the compound is provided in the manufacture of a medicament in any method of the present invention.

Figure 1 shows the dose response curve of a representative compound of the present invention ^{in the GLOSENSOR TM} analysis described in Biological Example 1. It should be noted that certain compounds are partial inhibitors, and the maximum inhibition of MALT1 para-apoptotic protease activity is about 70 to 80%.

Figure 2 shows the dose-response curve of a representative compound of the present invention in the IL-2 production analysis described in Biological Example 2. It should be noted that certain compounds are partial inhibitors, and the maximum inhibition of MALT1 para-apoptotic protease activity is about 50 to 80%.

1. Summary

This article describes compounds of formula (I), (II-A), or (II-B) that inhibit the activity of MALT1, and the compounds of the examples (herein collectively referred to as "compounds of the present invention"), and pharmaceutically acceptable compounds thereof salt. It also discloses a method of using it to inhibit MALT1 activity to treat diseases or symptoms including autoimmune disorders, inflammatory diseases, or cancers as described herein.

2. Definition

Unless otherwise indicated in this article, or clearly shielded from the context, "a(一)", "an(一)", "the (the)" and the context of the present invention (especially the context of the scope of patent application) as used herein Similar terms used in are intended to be understood to cover both the singular and the plural.

The term "halo" as used herein means halogen and includes chlorine, fluorine, bromine and iodine.

The term "alkyl" used alone or as part of a larger moiety, such as "alkoxy" or "haloalkyl", etc., means a saturated aliphatic linear or branched monovalent hydrocarbon group. Unless otherwise specified, the alkyl group generally has 1 to 4 or 1 to 6 carbon atoms, that is, (C ₁ -C ₄ )alkyl or (C ₁ -C ₆ )alkyl. Here, "(C ₁ -C ₄ )alkyl" means a group having 1 to 4 carbon atoms in a linear or branched arrangement; examples thereof include methyl, ethyl, n-propyl, isopropyl, N-butyl, secondary butyl, isobutyl, tertiary butyl, n-pentyl, isopentyl, neopentyl, n-hexyl, etc.

The term "C _1-6 alkylene" refers to a bivalent fully saturated branched or straight chain monovalent hydrocarbon group having 1 to 6 carbon atoms. Similarly, "C _1-4 alkylene", "C _1-3 alkylene" and "C _1-2 alkylene" are explained accordingly. Representative examples of C _1-6 alkylene include, but are not limited to, methylene, ethylidene, n-propylidene, isopropylidene, n-butylene, dibutylene, isobutylene, and tributylene. Butyl, n-pentyl, isopentyl, neopentyl, and n-hexyl.

"Optionally substituted hydroxy of C ₁ -C ₆ alkyl" means the above definition of C ₁ -C ₆ alkyl group which may be substituted by one or more hydroxyl groups. Examples include, but are not limited to, hydroxymethyl, hydroxyethyl, 1,2-dihydroxyethyl, 2,3-dihydroxy-propyl and the like.

As used herein, the term "di-C _1-6 alkylamino group" refers to _{a part of the formula -N(R a} )-R a, wherein each Ra is _{a C 1-6} alkyl group as defined above, which may be the same or different Analogously, "mono-C _1-6 alkylamino" refers to the formula -N (H) -R _a part of, wherein R _a is as hereinbefore defined under C _1-6 alkyl.

The term "alkenyl" means a branched or straight chain monovalent hydrocarbon group containing at least one double bond. Alkenyl groups can be monounsaturated or polyunsaturated, and can exist in the E or Z configuration. Unless otherwise specified, an alkenyl group generally has 2 to 6 carbon atoms, that is, a (C ₂ -C ₆ )alkenyl group. For example, "(C ₂ -C ₆ )alkenyl" means a group having 2 to 6 carbon atoms in a linear or branched arrangement.

The term "alkynyl" means a branched or straight chain monovalent hydrocarbon group containing at least one parametric bond. Unless otherwise specified, an alkynyl group generally has 2 to 6 carbon atoms, that is, a (C ₂ -C ₆ )alkynyl group. For example, "(C ₂ -C ₆ )alkynyl" means a group having 2 to 6 carbon atoms in a linear or branched arrangement.

The term "alkoxy" means an alkyl group connected through an oxygen linking atom, represented by -O-alkyl. For example, "C ₁ -C ₆ alkoxy" refers to -OC ₁ -C ₆ alkyl, where alkyl is defined herein, and "(C ₁ -C ₄ )alkoxy" includes methoxy, ethoxy Group, propoxy, and butoxy, etc. Representative examples of alkoxy include, but are not limited to, methoxy, ethoxy, 2-propoxy, butoxy, tertiary butoxy, pentoxy, hexyloxy, cyclopropoxy- , Cyclohexyloxy-etc. Generally, the alkoxy group has about 1 to 6 carbon atoms, 1 to 4 carbon atoms, or 1 to 2 carbon atoms.

The terms "haloalkyl" and "haloalkoxy", as the case may be, mean an alkyl or alkoxy group substituted with one or more halogen atoms. Examples of haloalkyl include, but are not limited to, trifluoromethyl, trichloromethyl, pentafluoroethyl and the like.

Therefore, the term "C _{1-6 alkyl substituted with halogen as required" refers to C 1} -C ₆ alkyl as defined above which can be substituted with one or more halogens. Examples include, but are not limited to, trifluoromethyl, difluoromethyl, fluoromethyl, trichloromethyl, 2,2,2-trifluoroethyl, 1-fluoromethyl-2-fluoroethyl, 3 -Bromo-2-fluoropropyl, and 1-bromomethyl-2-bromoethyl.

The term "cycloalkyl" as used herein includes saturated cyclic, bicyclic, tricyclic, or polycyclic hydrocarbon groups with 3 to 14 carbons containing the indicated number of rings and carbon atoms (e.g., C ₃ -C ₁₄ single Cyclic, C ₄ -C ₁₄ bicyclic, C ₅ -C ₁₄ tricyclic, or C ₆ -C ₁₄ polycyclic cycloalkyl). In some specific examples, "cycloalkyl" is a monocyclic cycloalkyl. Examples of monocyclic cycloalkyl groups include cyclopentyl (C ₅ ), cyclohexyl (C ₆ ), cyclopropyl (C ₃ ), cyclobutyl (C ₄ ), cycloheptyl (C ₇ ), and cyclooctyl (C ₈ ). In some specific examples, "cycloalkyl" is bicyclic cycloalkyl. Examples of bicyclic cycloalkyl groups include bicyclo[1.1.0]butane (C ₄ ), bicyclo[1.1.1]pentane (C ₅ ), spiro[2.2]pentane (C ₅ ), bicyclo[2.1.0] Pentane (C ₅ ), bicyclo[2.1.1] hexane (C ₆ ), bicyclo[3.3.3] undecane (C ₁₁ ), decalin (C ₁₀ ), bicyclo[4.3.2] eleven Alkane (C ₁₁ ), spiro[5.5]undecane (C ₁₁ ), and bicyclo[4.3.3]dodecane (C ₁₂ ). In some specific examples, "cycloalkyl" is tricyclic cycloalkyl. Examples of tricyclic cycloalkyl groups include adamantane (C ₁₂ ). Unless otherwise specified, "cycloalkyl" has three to six carbon atoms and is a monocyclic ring.

The term "aryl" used alone or as part of a larger group as in "aralkyl", "aralkoxy", or "aryloxyalkyl" means a carbocyclic aromatic ring. The term "aryl" can be used interchangeably with "aryl ring", "carbocyclic aromatic ring", "aryl" and "carbocyclic aryl". Generally, the aryl group is a monocyclic, bicyclic or tricyclic aryl group having 6 to 20 carbon atoms, usually 6 to 14 ring carbon atoms. Furthermore, the term "aryl" as used herein refers to an aromatic substituent which can be a single aromatic ring or multiple aromatic rings fused together. Examples include phenyl, naphthyl, anthracenyl, 1,2-dihydronaphthyl, 1,2,3,4-tetrahydronaphthyl, stilbyl, dihydroindenyl, indenyl and the like.

The "substituted aryl group" is substituted on any one or more substitutable ring atoms which are ring carbon atoms bonded to hydrogen. The substituted aryl group is usually substituted by 1 to 5 (for example, one, or two, or three) substituents, which are independently selected from the following groups: hydroxyl, thiol, cyano, nitro , C ₁ -C ₄ alkyl, C ₁ -C ₄ alkenyl, C ₁ -C ₄ alkynyl, C ₁ -C ₄ alkoxy, C ₁ -C ₄ thioalkyl, C ₁ -C ₄ alkenyl Oxy, C ₁ -C ₄ alkynyloxy, halogen, C ₁ -C ₄ alkylcarbonyl, carboxy, C ₁ -C ₄ alkoxycarbonyl, amine, C ₁ -C ₄ alkylamino, di-C _1- C ₄ alkylamino group, C ₁ -C ₄ alkyl amine carbonyl group, di-C ₁ -C ₄ alkyl amine carbonyl group, C ₁ -C ₄ alkyl carbonyl amine group, C ₁ -C ₄ alkyl carbonyl group, C ₁ -C ₄ alkylamino group, sulfonamide group, sulfamsulfonyl group, alkyl sulfonamide group, and C ₁ -C ₄ alkylaminosulfonyl group; wherein each of the foregoing hydrocarbon groups (e.g., alkyl, Alkenyl, alkynyl, alkoxy residues) may be further substituted with one or more residues independently selected from halogen, hydroxy or C ₁ -C ₄ alkoxy at each occurrence.

烷、嗎啉、1,4-二硫環己烷、哌

、哌啶、1,3-二氧雜環戊烷、咪唑啶、咪唑啉、吡咯啉、吡咯啶、四氫吡喃、二氫吡喃、氧硫雜環戊烷、二硫雜環戊烷、1,3-二

烷、1,3-二硫環己烷、氧硫雜環己烷、硫嗎啉等。 Words such as "heterocyclic group" or "heterocyclic group" mean a single ring, a non-aromatic (including partially saturated) ring with 3 to 10 members containing preferably 1 to 4 ring heteroatoms, or a preferably A polycyclic ring with 7 to 20 ring members and preferably 1 to 4 ring heteroatoms, wherein the polycyclic ring has one or more monocyclic non-aromatic heterocycles fused with one or more aromatic or heteroaromatic rings ring. Heterocyclyl groups usually have 3 to 7, 3 to 24, 4 to 16, 5 to 10, or 5 or 6 ring atoms; of which one to four, especially one or two ring atoms are heteroatoms (hence The remaining ring atoms are carbon). Each heteroatom is independently selected from nitrogen, quaternary nitrogen, nitrogen oxides (eg, NO); oxygen; and sulfur, including clumps and clumps. The heterocyclic group can be attached to a hetero atom or a carbon atom. Examples of heterocycles include tetrahydrofuran (THF), dihydrofuran, 1,4-dihydrofuran

Alkyl, morpholine, 1,4-disulfide cyclohexane, piperazine

, Piperidine, 1,3-dioxolane, imidazoline, imidazoline, pyrroline, pyrrolidine, tetrahydropyran, dihydropyran, oxathiolane, dithiolane , 1,3-two

Alkanes, 1,3-dithiocyclohexane, oxathane, thiomorpholine, etc.

Heterocyclic groups may include fused or bridged rings as well as spiro rings. In a specific example, the heterocyclic group is a bicyclic ring having a monocyclic non-aromatic heterocyclic ring fused with a phenyl group. Exemplary polycyclic heterocyclic groups include tetrahydroisoquinolinyl (e.g., 1,2,3,4-tetrahydroisoquinolin-7-yl, 2-methyl-1,2,3,4-tetrahydroisoquinolinyl Quinolin-7-yl, 1,2,3,4-tetrahydroisoquinolin-6-yl and 2-methyl-1,2,3,4-tetrahydroisoquinolin-6-yl), iso Indolinyl (e.g. 2-ethylisoindolin-5-yl, 2-methylisoindolin-5-yl), indolinyl, tetrahydrobenzo[f]oxazepin Oxazepinyl (for example, 2,3,4,5-tetrahydrobenzo[f][1,4]oxazepin-7-yl).

The words "heterocycle", "heterocyclic group", and "heterocyclic group", whether saturated or partially unsaturated, refer to rings that are optionally substituted. The substituted heterocyclic group may be a heterocyclic ring independently substituted with 1 to 4, for example, one, or two, or three or four substituents.

In some specific examples, the heterocyclyl group is a 3 to 14 membered non-aromatic ring system having ring carbon atoms and 1 to 4 ring heteroatoms, wherein each heteroatom is independently selected from nitrogen, oxygen, and sulfur ( "3- to 14-membered heterocyclic group").

Words such as "heteroaryl", "heteroaromatic", "heteroaryl ring", "heteroaryl group", "heteroaromatic ring" and "heteroaromatic group", used alone or as a larger part Part of the use, such as in "heteroaralkyl" or "heteroarylalkoxy", means having 5 to 14 carbons and at least one (usually 1 to 4, more usually 1 or 2 ) Aromatic ring groups of ring atoms of heteroatoms (for example, oxygen, nitrogen or sulfur). "Heteroaryl" includes The monocyclic heteroaromatic ring is fused to one or more other carbocyclic aromatic or heteroaromatic rings, monocyclic and polycyclic (for example, bicyclic or tricyclic). Therefore, "5- to 14-membered heteroaryl" includes monocyclic, bicyclic or tricyclic ring systems.

唑基(例如，3-異

唑基、4-異

唑基、5-異

唑基)、

二唑基、(例如，2-

二唑基、5-

二唑基)、

唑基(例如，2-

唑基、4-

唑基、5-

唑基)、吡唑基(例如，3-吡唑基、4-吡唑基)、吡咯基(例如，1-吡咯基、2-吡咯基、3-吡咯基)、吡啶基(例如，2-吡啶基、3-吡啶基、4-吡啶基)、嘧啶基(例如，2-嘧啶基、4-嘧啶基、5-嘧啶基)、嗒

基(例如，3-嗒

基)、噻唑基(例如，2-噻唑基、4-噻唑基、5-噻唑基)、三唑基(例如，2-三唑基、5-三唑基)、四唑基(例如，四唑基)、噻吩基(例如，2-噻吩基、3-噻吩基)、嘧啶基、吡啶基與嗒

基。 Examples of 5- to 6-membered monocyclic heteroaryl groups include furyl (e.g., 2-furyl, 3-furyl), imidazolyl (e.g., N -imidazolyl, 2-imidazolyl, 4-imidazolyl, 5- Imidazolyl), iso

Azolyl (e.g., 3-iso

Azolyl, 4-iso

Azolyl, 5-iso

Azole),

Diazolyl, (e.g. 2-

Diazolyl, 5-

Diazolyl),

Azolyl (e.g. 2-

Azolyl, 4-

Azolyl, 5-

Azolyl), pyrazolyl (e.g., 3-pyrazolyl, 4-pyrazolyl), pyrrolyl (e.g., 1-pyrrolyl, 2-pyrrolyl, 3-pyrrolyl), pyridyl (e.g., 2 -Pyridyl, 3-pyridyl, 4-pyridyl), pyrimidinyl (e.g., 2-pyrimidinyl, 4-pyrimidinyl, 5-pyrimidinyl),

Base (for example, 3-ta

Group), thiazolyl (e.g., 2-thiazolyl, 4-thiazolyl, 5-thiazolyl), triazolyl (e.g., 2-triazolyl, 5-triazolyl), tetrazolyl (e.g., four Azolyl), thienyl (e.g., 2-thienyl, 3-thienyl), pyrimidinyl, pyridyl, and

base.

唑基、3-、4-、或5-異

唑基、3-或5-1,2,4-三唑基、4-或5-1,2,3-三唑基、四唑基、2-、3-、或4-吡啶基、3-或4-嗒

基、3-、4-、或5-吡

基、2-吡

基、與2-、4-、或5-嘧啶基。 Typically, the heteroaryl group is a 5 to 10 membered ring system (e.g., a 5 to 6 membered monocyclic ring or an 8 to 10 membered bicyclic ring) or a 5 to 6 membered ring system. Typical heteroaryl groups include 2- or 3-thienyl, 2- or 3-furyl, 2- or 3-pyrrolyl, 2-, 4-, or 5-imidazolyl, 3-, 4-, or 5 -Pyrazolyl, 2-, 4-, or 5-thiazolyl, 3-, 4-, or 5-isothiazolyl, 2-, 4-, or 5-

Azolyl, 3-, 4-, or 5-iso

Azolyl, 3- or 5-1,2,4-triazolyl, 4- or 5-1,2,3-triazolyl, tetrazolyl, 2-, 3-, or 4-pyridyl, 3 -Or 4-ta

Base, 3-, 4-, or 5-pyridine

Base, 2-pyridine

Group, and 2-, 4-, or 5-pyrimidinyl.

唑基、苯并咪唑基、異喹啉基、吲哚基、異吲哚基、吖啶基、或苯并異

唑基。 Examples of polycyclic aromatic heteroaryl groups include carbazolyl, benzimidazolyl, benzothienyl, benzofuranyl, indolyl, quinolinyl, benzotriazolyl, benzothiazolyl, benzo

Azolyl, benzimidazolyl, isoquinolinyl, indolyl, isoindolyl, acridinyl, or benziso

Azole.

基、2-、3-、4-、5-、或6-萘啶基、2-、3-、5-、6-、7-、或8-喹唑啉基、3-、4-、5-、6-、7-、或8-噌啉基、2-、4-、6-、或7-喋啶基、1-、2-、3-、4-、5-、6-、7-、或8-4aH咔唑基、1-、2-、3-、4-、5-、6-、7-、或8-咔唑基、1-、3-、4-、5-、6-、7-、8-、或9-咔啉基、1-、2-、3-、4-、6-、7-、8-、9-、或10-啡啶基、1-、2-、3-、4-、5-、6-、7-、8-、或9-吖啶基、1-、2-、4-、5-、6-、7-、8-、或9-苯并喹唑啉基(perimidinyl)、2-、3-、4-、5-、6--8-、9-、或10-啡咯啉基、1-、2-、3-、4-、6-、7-、8-、或9-啡

基、1-、2-、3-、4-、6-、7-、8-、9-、或10-啡噻

基、1-、2-、3-、4-、6-、7-、8-、9-、或10-啡

基、2-、3-、4-、5-、6-、或1-、3-、4-、5-、6-、7-、8-、9-、或10-苯并異喹啉基、2-、3-、4-、或噻吩并[2,3-b]呋喃基、2-、3-、5-、6-、7-、8-、9-、10-、或11-7H-吡

并[2,3-c]咔唑基、2-、3-、5-、6-、或7-2H-呋喃并[3,2-b]-吡喃基、2-、3-、4-、5-、7-、或8-5H-吡啶并[2,3-d]-o-

基、1-、3-、或5-1H-吡唑并[4,3-d]-

唑基、2-、4-、或5-4H-咪唑并[4,5-d]-噻唑基、3-、5-、或8-吡

并[2,3-d]嗒

基、2-、3-、5-、或6-咪唑并[2,1-b]-噻唑基、1-、3-、6-、7-、8-、或9-呋喃并[3,4-c]噌啉基、1-、2-、3-、4-、5-、6-、8-、9-、10-、或11-4H-吡啶并[2,3-c]咔唑基、2-、3-、8-、或7-咪唑并[1,2-b][1,2,4]三

基、7-苯并[b]噻吩基、2-、4-、5-、6-、或7-苯并

唑基、2-、4-、5-、6-、或7-苯并咪唑基、2-、4-、4-、5-、6-、或7-苯并噻唑基、1-、2-、4-、5-、6-、7-、8-、或9-苯并奧沙平基(benzoxapinyl)、2-、4-、 5-、6-、7-、或8-苯并

基(benzoxazinyl)、1-、2-、3-、5-、6-、7-、8-、9-、10-、或11-1H-吡咯并[1,2-b][2]苯扎氮平基(benzazapinyl)。 Therefore, the term "heteroaryl" also refers to a group in which a heteroaromatic ring is fused with one or more aryl, alicyclic, or heterocyclic ring, wherein the group or point of attachment is in the heterocyclic ring. On the aromatic ring. Non-limiting examples include 1-, 2-, 3-, 5-, 6-, 7-, or 8-indazinyl, 1-, 3-, 4-, 5-, 6-, or 7-isoindole Base, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, or 7-indazolyl, 2-, 4 -, 5-, 6-, 7-, or 8-purinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, or 9-quinazinyl, 2-, 3- , 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6-, 7-, or 8-isoquinolinyl, 1-, 4 -, 5-, 6-, 7-, or 8-phthalein

Group, 2-, 3-, 4-, 5-, or 6-naphthyridinyl, 2-, 3-, 5-, 6-, 7-, or 8-quinazolinyl, 3-, 4-, 5-, 6-, 7-, or 8-cinnolinyl, 2-, 4-, 6-, or 7-pteridinyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-4aH carbazolyl, 1-, 2-, 3-, 4-, 5-, 6-, 7-, or 8-carbazolyl, 1-, 3-, 4-, 5- , 6-, 7-, 8-, or 9-carbolinyl, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenanthridinyl, 1- , 2-, 3-, 4-, 5-, 6-, 7-, 8-, or 9-acridinyl, 1-, 2-, 4-, 5-, 6-, 7-, 8-, Or 9-benzoquinazolinyl (perimidinyl), 2-, 3-, 4-, 5-, 6--8-, 9-, or 10-phenanthrolinyl, 1-, 2-, 3- , 4-, 6-, 7-, 8-, or 9-brown

Base, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-phenothi

Base, 1-, 2-, 3-, 4-, 6-, 7-, 8-, 9-, or 10-brown

Base, 2-, 3-, 4-, 5-, 6-, or 1-, 3-, 4-, 5-, 6-, 7-, 8-, 9-, or 10-benzisoquinoline Group, 2-, 3-, 4-, or thieno[2,3- b ]furyl, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11 -7H-pyridine

And [2,3-c]carbazolyl, 2-, 3-, 5-, 6-, or 7-2H-furo[3,2-b]-pyranyl, 2-, 3-, 4 -, 5-, 7-, or 8-5H-pyrido[2,3-d]-o-

Group, 1-, 3-, or 5-1H-pyrazolo[4,3-d]-

Azolyl, 2-, 4-, or 5-4H-imidazo[4,5-d]-thiazolyl, 3-, 5-, or 8-pyridine

And [2,3-d] click

Group, 2-, 3-, 5-, or 6-imidazo[2,1-b]-thiazolyl, 1-, 3-, 6-, 7-, 8-, or 9-furo[3, 4-c] Cinolinyl, 1-, 2-, 3-, 4-, 5-, 6-, 8-, 9-, 10-, or 11-4H-pyrido[2,3-c]carba Azolyl, 2-, 3-, 8-, or 7-imidazo[1,2-b][1,2,4] three

Group, 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzo

Azolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, 2-, 4-, 4-, 5-, 6-, or 7-benzothiazolyl, 1-, 2 -, 4-, 5-, 6-, 7-, 8-, or 9-benzoxapinyl, 2-, 4-, 5-, 6-, 7-, or 8-benzoxapinyl

Benzoxazinyl, 1-, 2-, 3-, 5-, 6-, 7-, 8-, 9-, 10-, or 11-1H-pyrrolo[1,2-b][2]benzene Zalazapinyl (benzazapinyl).

唑基、2-、4-、5-、6-、或7-苯并咪唑基、與2-、4-、5-、6-、或7-苯并噻唑基。 Typical fused heteroaryl groups include, but are not limited to 2-, 3-, 4-, 5-, 6-, 7-, or 8-quinolinyl, 1-, 3-, 4-, 5-, 6 -, 7-, or 8-isoquinolinyl, 2-, 3-, 4-, 5-, 6-, or 7-indolyl, 2-, 3-, 4-, 5-, 6-, Or 7-benzo[b]thienyl, 2-, 4-, 5-, 6-, or 7-benzo

Azolyl, 2-, 4-, 5-, 6-, or 7-benzimidazolyl, and 2-, 4-, 5-, 6-, or 7-benzothiazolyl.

The term pyridine or pyridyl optionally substituted with hydroxy as used herein, for example 2-pyridyl, 3-pyridyl, or 4-pyridyl refers to the respective hydroxypyridine or hydroxy-pyridyl, and may include their mutual variations Configurations such as pyridone or pyridone-yi respectively.

As used herein, the term pyridine or pyridyl optionally substituted with pendant oxy groups, such as 2-pyridyl, 3-pyridyl, or 4-pyridyl, refers to the respective pyridone or pyridone group and may include their interactions. Tautomers, such as hydroxy-pyridine or hydroxy-pyridyl, respectively, as long as the tautomers are available. If necessary, the pyridine or pyridyl group substituted by the pendant oxy group can be further referred to the respective pyridine-N-oxide or pyridyl-N-oxide.

"Substituted heteroaryl" is substituted on any one or more substitutable ring atoms which are ring carbon or ring nitrogen atoms bonded to hydrogen.

The term "bridged bicyclic group" refers to a ring system that includes two rings that share at least three adjacent ring atoms.

As used herein, many moieties (eg, alkyl, alkylene, cycloalkyl, aryl, heteroaryl, or heterocyclyl) are referred to as "substituted" or "optionally substituted." When a part is modified by one of these terms, unless otherwise specified, it means that the part is known to those skilled in the art to which the invention pertains to be used for substitution. Any part can be substituted, including one or more (e.g., one to Three) substituents. If it exists If there is more than one substituent, each substituent can be independently selected. Such substitution methods are known in the technical field of the invention and/or taught by the present invention. The optional substituent may be any substituent suitable for attaching the moiety.

In the case of not specifically enumerating the appropriate substituents, the exemplified substituents include, but are not limited to: (C ₁ -C ₅ )alkyl, (C ₁ -C ₅ )hydroxyalkyl, (C ₁ -C ₅ )haloalkane Group, (C ₁ -C ₅ )alkoxy, (C ₁ -C ₅ )haloalkoxy, halogen, hydroxyl, cyano, amino, -CN, -NO ₂ , -OR ^c1 , -NR ^a1 R ^b1 , -S(O) _i R ^a1 , -NR ^a1 S(O) _i R ^b1 , -S(O) _i NR ^a1 R ^b1 , -C(=O)OR ^a1 , -OC(=O)OR ^a1 , -C(=S)OR ^a1 , -O(C=S)R ^a1 , -C(=O)NR ^a1 R ^b1 , -NR ^a1 C(=O)R ^b1 , -C(=S)NR ^a1 R ^b1 , -C(=O)R ^a1 , -C(=S)R ^a1 , NR ^a1 C(=S)R ^b1 , -O(C=O)NR ^a1 R ^b1 , -NR ^a1 (C=S )OR ^b1 , -O(C=S)NR ^a1 R ^b1 , -NR ^a1 (C=O)NR ^a1 R ^b1 , NR ^a1 (C=S)NR ^a1 R ^b1 , phenyl, or 5 to 6 members Aryl. Each R ^a1 and each R ^{b1 are} independently selected from -H and (C ₁ -C ₅ ) alkyl, optionally substituted by hydroxy or (C ₁ -C ₃ ) alkoxy; R ^c1 is -H, (C ₁ -C ₅ )haloalkyl or (C ₁ -C ₅ )alkyl, wherein (C ₁ -C ₅ )alkyl is optionally substituted with hydroxy or (C ₁ -C _{3 )alkoxy.}

The compounds described herein can exist in various tautomeric forms. Words such as "tautomers" or "tautomerism" refer to changes in at least one form and at least one valence of a hydrogen atom (for example, a single bond becomes a double bond, a parametric bond becomes a single bond, or vice versa ) Two or more interconvertible compounds/substitutes produced. Exemplary tautomerism includes, ketone to enol, amide to amide, internal amide to internal amide, enamine to imine, and enamine to (different enamine) tautomerism. This teaching covers compounds in the form of tautomers, including forms that are not structurally depicted. All such isomeric forms of such compounds are expressly included. If the tautomer of a compound is aromatic, the compound is aromatic.

It should be understood that when the compound herein is represented by a structural formula or is represented by a chemical name herein Obviously, all other tautomeric forms that the compound may exist are covered by the structural formula.

Compounds of any of the above formulas may exhibit one or more isomerism (for example, optical, geometric, or tautomerism). Compounds of any of the above formulas can also be isotopically labeled. Since the compounds of any of the above formulas are defined by referring to their structural features, such variations are implied therein, and therefore fall within the scope of this disclosure.

The compounds of any of the above formulas containing one or more asymmetric carbon atoms may exist as two or more stereoisomers. In the case where the compound of any of the above formulas contains alkenyl or alkenylene, geometric cis/trans (or Z/E) isomers may exist. When structural isomers are converted into each other through a low-energy barrier, tautomerism ("tautomerism") occurs. Compounds of any of the above formulas containing, for example, imino groups, ketone groups, or oxime groups can take the form of proton tautomerism, or so-called valence tautomerism in compounds containing aromatic groups. Therefore, a single compound may exhibit more than one type of isomerism.

Compounds with one or more palm centers can exist in a variety of stereoisomeric forms. Stereoisomers are compounds that differ only in their spatial arrangement. Stereoisomers include all diastereoisomers, enantiomers, and epimeric forms, racemates and mixtures thereof. The term "geometric isomer" refers to a compound having at least one double bond, wherein the double bond can be in the cis form [also known as the same (syn) or entgegen ( E )] or trans [also known as the anti (anti ) Or together (zusammen, Z )] forms and mixtures thereof. When the disclosed compound is named or depicted by structure without indicating stereochemistry, it is generally understood that the name or structure covers one or more possible stereoisomers, or geometric isomers, or the covered stereoisomers or A mixture of geometric isomers.

When describing geometric isomers by name or structure, it should be understood that the named or described isomer exists to a higher degree than the other isomer, that is, the geometric isomer named or described a few The purity of any isomer is greater than 50%, for example at least 60%, 70%, 80%, 90%, 99%, or 99.9% by weight. The purity of geometric isomers is determined by dividing the weight of the named or depicted geometric isomers in the mixture by the total weight of all geometric isomers in the mixture.

A racemic mixture means 50% of an enantiomer and 50% of the corresponding enantiomer. When naming or describing a compound with a palm center without indicating the stereochemistry of the palm center, it is generally understood that the name or structure encompasses the two possible enantiomer forms of the compound (for example, the pure enantiomer , Enrichment or racemization of mirror image isomers). When naming or describing a compound with two or more palm centers without indicating the stereochemistry of the palm center, it is generally understood that the name or structure encompasses all possible diastereomer forms of the compound [for example, non- Pure enantiomers, enrichment of diastereoisomers, and equimolar mixtures of one or more diastereoisomers (for example, racemic mixtures)].

The mixture of enantiomers and diastereomers can be separated into constituent enantiomers or stereoisomers by commonly known methods, such as palm phase gas chromatography and palm phase high performance liquid chromatography , The compound is crystallized into a palm salt complex; or the compound is crystallized in a palm solvent. It is also possible to use commonly known asymmetric synthesis methods to obtain the enantiomers and diastereomers with diastereomers or pure intermediates, reagents, and catalysts of the enantiomers.

When a compound is designated by the name or structure of a single enantiomer, unless otherwise indicated, the optical purity of the compound is at least 60%, 70%, 80%, 90%, 99% or 99.9% (also Referred to as "mirror isomers pure"). Optical purity is the weight of the named or depicted enantiomer in the mixture divided by the total weight of the two enantiomers in the mixture.

When the stereochemistry of the disclosed compound is named or depicted by a structure, and the named or depicted structure encompasses more than one stereoisomer (e.g., as in a diastereomer pair), it should be understood that Covering one of the stereoisomers or any mixture of covered stereoisomers is also included. should It is further understood that the stereoisomeric purity of the named or depicted stereoisomer is at least 60%, 70%, 80%, 90%, 99% or 99.9% by weight. In this case, the purity of the stereoisomers is determined by dividing the total weight of the stereoisomers covered by the name or structure in the mixture by the total weight of all stereoisomers in the mixture.

The pharmaceutically acceptable salt of the compound of any of the above formulas may also contain opposing ions with optical activity (such as d-lactic acid or 1-lysine acid) or racemic (such as dl-tartaric acid or dl-arginine) .

The cis/trans isomers can be separated using well-known techniques known to those skilled in the art to which the invention pertains, for example, chromatography and segmented crystallization.

Conventional techniques for preparing/isolating individual enantiomers include palm synthesis from appropriate optically pure precursors, or isolation of racemates (or salts) using, for example, palm high pressure liquid chromatography (HPLC) Or derivatives of racemates). Alternatively, the racemate (or racemic precursor) can be reacted with a suitable optically active compound (for example, alcohol), or in the case of a compound containing acidic or basic groups of any of the above formulas, React with base or acid (for example 1-phenylethylamine or tartaric acid). The resulting mixture of diastereomers can be separated by chromatography and/or segmented crystallization, and then one or two diastereomers can be converted into the corresponding pure enantiomers by means known to the skilled person. Things. The palm compounds of any of the above formulas (and their palm precursors) can be obtained in the form of enrichment of enantiomers using chromatography. The chromatography is generally HPLC, using asymmetric resins and hydrocarbons. (Generally heptane or hexane) consisting of a mobile phase, which contains 0 to 50% by volume of isopropanol, generally 2% to 20%, and 0 to 5% by volume of alkylamine, generally 0.1% Ethylamine. The eluent is concentrated to obtain an enriched mixture. The palm chromatography method using subcritical and supercritical fluids can be used. In some specific examples of the present disclosure, the method for palm chromatographic analysis is known in the technical field of the invention [see, for example, Smith, Roger M., Loughborough University, Loughborough, UK; Chromatographic Science Series (1998), 75 (Supercritical Fluid Chromatography with Packed Columns), pp.223-249 and references cited therein]. The column is available from Chiral Technologies, Inc, West Chester, Pa., USA, which is a subsidiary of Daicel® Chemical Industries, Ltd., Tokyo, Japan.

It must be emphasized that the compound of any of the above formulas has been formed as a single tautomer herein, and all possible tautomers are within the scope of this disclosure.

The present disclosure also includes all pharmaceutically acceptable isotope-labeled compounds of any of the above-mentioned formulas, in which one or more atoms have the same atomic number, but the atomic weight or mass number is different from the atomic weight or mass number that is the majority in nature Atomic substitution.

Examples of isotopes suitable for inclusion in the compounds of the present disclosure include hydrogen (e.g. ² H and ³ H), carbon (e.g. ¹¹ C, ¹³ C and ¹⁴ C), chlorine (e.g. ³⁶ Cl), fluorine (e.g. ¹⁸ F), Iodine (such as ¹²³ I and ¹²⁵ I), nitrogen (such as ¹³ N and ¹⁵ N), oxygen (such as ¹⁵ O, ¹⁷ O and ¹⁸ O), phosphorus (such as ³² P) and sulfur (such as ³⁵ S) isotope.

Specific isotope-labeled compounds of any of the above formulas, for example, those incorporating radioisotopes, are useful in the study of drug and/or matrix tissue distribution. ^{For this purpose, the radioisotopes tritium (i.e. 3} H) and carbon-14 (i.e. ¹⁴ C) are particularly useful in view of their easy to incorporate and readily available detection methods.

Substitution with heavier isotopes such as deuterium (ie ² H) provides specific therapeutic advantages due to greater metabolic stability, for example, increased in vivo half-life or reduced dosage requirements.

Replacement with positron emission isotopes (such as ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N) can be used in positron emission tomography (PET) studies to check the position occupancy of the matrix receptor.

The isotope-labeled compound of any of the above-mentioned formulas can usually use the well-known techniques of those skilled in the art to which the invention pertains, or use appropriate isotope-labeled methods similar to those described in the accompanying examples and preparations. The reagents are prepared instead of the previously used non-labeled reagents.

The pharmaceutically acceptable solvates according to the present disclosure include those in which the crystallization solvent can be replaced by isotope, for example, D ₂ O, d ₆ -acetone, and d ₆ -DMSO.

The formula given herein is also intended to represent any unlabeled forms as well as isotopic forms of the compounds of the marker, e.g. ² H. Furthermore, it is generally understood that in this context, deuterium is regarded as a substituent of the compound of formula (I), which is replaced by deuterium (ie ² H or D) due to greater metabolic stability (for example, increased in vivo half-life or Reducing the dose requirement or improving the therapeutic index) can provide specific therapeutic advantages. The concentration of these heavier isotopes (specifically deuterium) can be defined by the isotope enrichment factor. The term "isotopic enrichment factor" as used herein means the ratio between the isotopic abundance of a specified isotope and the natural abundance. If the substituent in the compound of the present invention is indicated as deuterium, the isotope enrichment factor of this compound for each designated deuterium atom is at least 3500 (52.5% deuterium incorporation for each designated deuterium atom), at least 4000 (60% deuterium incorporation) ), at least 4500 (67.5% deuterium incorporated), at least 5000 (75% deuterium incorporated), at least 5500 (82.5% deuterium incorporated), at least 6000 (90% deuterium incorporated), at least 6333.3 (95% deuterium incorporated) ), at least 6466.7 (97% deuterium incorporated), at least 6600 (99% deuterium incorporated), or at least 6633.3 (99.5% deuterium incorporated).

The compound of the present invention may exist in a free form for treatment, or, where appropriate, in a pharmaceutically acceptable salt form.

The terms "salts" or "salts" as used herein refer to acid addition salts or base addition salts of the compounds of the present invention. "Salts" especially include "pharmaceutically acceptable salts". The term "pharmaceutically acceptable salt" refers to a salt that retains the biological efficacy and properties of the compound of the present invention and is usually not biologically or otherwise unfavorable; in many cases, the compound of the present invention can rely on amine groups and/ Or the presence of carboxyl or similar groups to form acid and/or alkali salts.

Pharmaceutically acceptable acid addition salts can be formed with inorganic acids and organic acids, for example, acetate, aspartate, benzoate, sulfonate, bromide/hydrobromide, bicarbonate/carbonic acid Salt, sulfur Bisulphate/sulfate, camphorsulfonate, chloride/hydrochloride, chlortheophylionate, citrate, ethanedisulfonate, fumarate, glucoheptonate, gluconic acid Salt, glucuronate, hippurate, hydroiodide/iodide, 2-isethionate, lactate, lactobionate, lauryl sulfate, malate, maleic acid Salt, malonate, phenylglycolate, methanesulfonate, methylsulfate, naphthoate, naphthalenesulfonate, nicotinate, nitrate, octadecanoate, oleate, oxalic acid Salt, palmitate, hydroxynaphthate, phosphate/hydrogen phosphate/dihydrogen phosphate, polygalacturonate, propionate, stearate, succinate, hyposalicylate, tartaric acid Salt, tosylate and trifluoroacetate.

Inorganic acids from which salts can be derived include, for example, hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like.

Organic acids from which salts can be derived include, for example, acetic acid, propionic acid, glycolic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, mandelic acid , Methanesulfonic acid, ethanesulfonic acid, toluenesulfonic acid, salamic acid, etc. Pharmaceutically acceptable base addition salts can be formed with inorganic bases and organic bases. Inorganic bases from which salts can be derived include, for example, ammonium salts and metals in columns I to XII of the periodic table; in certain specific examples, the salts are derived from sodium, potassium, ammonium, calcium, magnesium, iron, silver, zinc and Copper; particularly suitable salts include ammonium, potassium, sodium, calcium and magnesium salts.

與三木甲胺(tromethamine)。 Organic bases from which salts can be derivatized include, for example, primary, secondary and tertiary amines, and substituted amines include naturally occurring substituted amines, cyclic amines, and basic ion exchange resins. Specific organic amines include isopropylamine, benzylethylenediamine, choline salt, diethanolamine, diethylamine, lysine, meglumine, piperidine

With tromethamine (tromethamine).

The pharmaceutically acceptable salts of the present invention can be synthesized with basic or acidic groups using conventional chemical methods. Generally, such salts can be reacted by reacting the free acid form of these compounds with a stoichiometric appropriate base (such as hydroxide, carbonate, bicarbonate, etc.) of Na, Ca, Mg or K, or by reacting these compounds The free base form of the compound is prepared by reacting with a stoichiometric amount of appropriate acid. Such reactions are generally carried out in water or organic solvents or a mixture of the two. Generally, where feasible, it is desirable to use a non-aqueous medium such as diethyl ether, ethyl acetate, ethanol, isopropanol, or acetonitrile. More lists of suitable salts can be found in, for example, "Remington's Pharmaceutical Sciences," 20th ed., Mack Publishing Company, Easton, PA, (1985); and "Handbook of Pharmaceutical Salts: Properties, Selection, and Use" by Stahl and Wermuth ( Wiley-VCH, Weinheim, Germany, 2002).

The terms "composition" and "composition" are used interchangeably.

The "subject" is a mammal, preferably a human, but can also be an animal that requires veterinary treatment, for example, companion animals (for example, dogs, cats, etc.), farm animals (for example, cows, sheep, pigs, horses, etc.) Etc.) and experimental animals (for example, rats, mice, guinea pigs, etc.).

As used herein, if a subject can derive biological, medical, or quality of life benefits from such treatment, then the subject "needs" treatment.

The terms "administer", "administering" and "administration" refer to methods of introducing the compound of the present invention or its composition into or on the body of a subject. These methods include, but are not limited to, intra-articular (in joints), intravenous, intramuscular, intratumor, intradermal, intraperitoneal, subcutaneous, oral, topical, spinal cord, inhalation, transdermal, rectal, etc. . Drug administration techniques that can be used with the formulations and methods described herein are described in, for example, Goodman and Gilman, The Pharmacological Basis of Therapeutics , current ed.; Pergamon; and Remington's, Pharmaceutical Sciences (current edition), Mack Publishing Co., Easton, Pennsylvania.

As used herein, the terms "inhibit", "inhibition" and "inhibit" refer to the reduction or suppression of a given symptom, sign, or disease, or disease, or the baseline of biological activity or process Significant reduction in activity.

"Treatment" and other words refer to reversing, alleviating, or inhibiting the progression of the diseases described herein. In some specific cases, treatment may be administered after one or more symptoms or signs of the disease have developed or have been observed (ie, therapeutic treatment). In other specific cases, the treatment can be administered in the absence of symptoms or signs of the disease. For example, treatment can be administered to susceptible subjects before the symptoms begin (ie, prophylactic treatment) (e.g., based on history of symptoms and/or based on exposure to pathogens). Treatment can also be continued after the symptoms have disappeared, for example, to delay or prevent recurrence.

The words "symptom", "disease", and "illness" are used interchangeably.

Generally, the effective amount of the compound taught herein depends on various factors, such as a given drug or compound, pharmaceutical formulation, route of administration, type of disease or condition, identity of the subject or host to be treated, etc., but it can still be achieved through familiarity. A person skilled in the technical field to which the invention belongs routinely decides. The effective amount of the compound of the present teaching can be easily determined by a person having ordinary knowledge in the technical field to which the invention belongs, and through routine methods known in the technical field to which the invention belongs.

The term "effective amount" means an amount that results in beneficial or desired results when administered to a subject, including clinical results, for example, when compared to a control group, the amount that inhibits, suppresses, or alleviates the symptoms of the subject being treated. For example, the effective amount can be administered in a unit dosage form (e.g., 1 mg to about 50 g per day, for example, 1 mg to about 5 g per day).

The term "therapeutically effective amount" of the compound of the present invention refers to the biological or medical response that will elicit the subject, for example, reducing or inhibiting the activity of enzymes or proteins, or improving symptoms, reducing symptoms, slowing down or delaying the progression of the disease, Or the amount of disease prevention. In a non-limiting specific example, the term "therapeutically effective amount" refers to the amount of the compound of the present invention, which, when administered to a subject, effectively (1) at least partially slows down, inhibits, prevents and/or improves (i) via MALT1 mediator, or (ii) related to MALT1 activity, or (iii) symptoms, or disorders or diseases characterized by MALT1 activity (normal or abnormal); or (2) reducing or inhibiting The activity of MALT1; or (3) reduce or inhibit the performance of MALT1; or (4) modify the protein quality of MALT1. In another non-limiting embodiment, the term "therapeutically effective amount" refers to the amount of the compound of the present invention, which when administered to cells, or tissues, or non-cellular biological materials, or media, is effective to at least partially reduce or Inhibit the activity of MALT1; or partially or completely reduce or inhibit the performance of MALT1.

Unless otherwise indicated herein or clearly conflicting with the context, all methods described herein can be performed in any appropriate order. The use of any and other examples or illustrative language (such as "for example") provided herein is only intended to better illustrate the present invention, and is not intended to limit the scope of the present invention.

The general chemical terms used in the above formula have their usual meanings.

As used herein, "h" or "hr" refers to hours, "min" refers to minutes, "MCL" refers to adventitia cell lymphoma, "AML" refers to acute myeloid leukemia, and "CML" refers to chronic type For myelogenous leukemia, "Boc" refers to N-tertiary butoxycarbonyl, "EA" refers to ethyl acetate, "DCM" refers to dichloromethane, "DMSO" refers to dimethyl sulfoxide, and "DMA" refers to Dimethylacetamide, "THF" refers to tetrahydrofuran, "MtBE" refers to methyl tertiary butyl ether, "TEA" refers to triethylamine, "FBS" refers to fetal bovine serum, and "PBS" refers to Phosphate buffered saline, "BSA" means bovine serum albumin, "RT" means room temperature, "mpk" means milligrams per kilogram, "po" means oral administration (oral), "qd" means It is administered once a day, "HPLC" means high pressure liquid chromatography, "q2d" means once every two days, "q2dx10" means 10 times a single dose every two days, and "VSMC" means vascular smooth muscle cells and "XRD" means X-ray diffraction.

The term "pharmaceutically acceptable carrier" as used herein includes any and all solvents, dispersion media, coatings, surfactants, antioxidants, preservatives (e.g., antioxidants) known to those skilled in the art to which the invention belongs. Bacterial agents, antifungal agents), isotonic agents, absorption retardants, salts, preservatives, drug stabilizers, binders, excipients, disintegrants, lubricants, sweeteners, flavoring agents, dyes, etc. Combinations (see, for example, Remington's Pharmaceutical Sciences, 18th Ed. Mack Printing Company, 1990, pp. 1289-1329). Unless any conventional carrier is incompatible with the active ingredient, use in therapeutic or pharmaceutical compositions will be considered.

Furthermore, the compounds of the present invention, including their salts, can also be obtained in the form of their hydrates, or contain other solvents used for their crystallization. The compounds of the present invention can form solvates inherently or by design with pharmaceutically acceptable solvents (including water); therefore, the present invention is intended to include both solvated and non-solvated forms. The term "solvate" refers to a molecular complex of the compound of the present invention (including its pharmaceutically acceptable salts) and one or more solvent molecules. Such solvent molecules are often used in the technical field of medical inventions and are known to be harmless to recipients, such as water, ethanol, etc. The term "hydrate" refers to a complex in which the solvent molecule is water.

The compounds of the present invention, including their salts, hydrates and solvates, can form polymorphs either inherently or by design. In another aspect, the present invention provides a pharmaceutical composition comprising a compound of the present invention and a pharmaceutically acceptable carrier. The pharmaceutical composition can be formulated for specific administration routes, such as oral administration, injection administration, and rectal administration. In addition, the pharmaceutical composition of the present invention can be composed in solid form (including but not limited to capsules, lozenges, pills, granules, powders or suppositories) or in liquid form (including but not limited to solutions, suspensions or emulsions). The pharmaceutical composition can be subjected to conventional medical operations, such as sterilization, and/or can contain conventional inert diluents, lubricants, or buffers, and adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers and buffers剂 etc.

Typically, the pharmaceutical composition is a lozenge or gelatin capsule, which contains the active ingredient and a) diluent, for example, lactose, dextrose, sucrose, mannitol, sorbitol, cellulose and/or glycine; b) Lubricants, such as silicon dioxide, talc, stearic acid, its magnesium or calcium salts and/or polyethylene glycol; and c) adhesives for lozenges, such as magnesium aluminum silicate, starch paste , Gelatin, tragacanth, methyl cellulose, Sodium carboxymethyl cellulose and/or polyvinylpyrrolidone; if necessary d) disintegrant, for example, starch, agar, alginic acid or its sodium salt, or foaming mixture; and/or e) absorbent, coloring agent , Flavoring and sweetening agents. The tablet may be coated with a film or enteric coating according to a method known in the technical field to which the invention belongs.

Suitable compositions for oral administration include effective amounts in the form of tablets, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsions, hard or soft capsules, or syrups or elixirs, etc. Compounds of the invention. According to any method known in the technical field of the invention for the manufacture of pharmaceutical compositions, a composition intended for oral use is prepared. Such composition may contain one or more selected from sweeteners, flavoring agents, coloring agents and preservatives. The medicaments that make up the group, so as to provide medicinal exquisite and tasteful preparations. Tablets may contain active ingredients incorporated with non-toxic pharmaceutically acceptable excipients suitable for the manufacture of tablets. These excipients are, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating agents and disintegrating agents, such as corn starch or alginic acid; binding agents, such as starch , Gelatin or gum arabic; and lubricants such as magnesium stearate, stearic acid or talc. Using known techniques to make the tablet uncoated or coated to delay its disintegration and absorption in the gastrointestinal tract, so as to provide a sustained effect over a longer period of time. For example, time delay materials such as glyceryl monostearate or glyceryl distearate can be used. Oral formulations can be presented in hard gelatin capsules, in which the active ingredient is mixed with an inert solid diluent (such as calcium carbonate, calcium phosphate or kaolin); or presented in soft gelatin capsules, in which the active ingredient is mixed with water or an oil medium (such as peanut oil, Liquid paraffin or olive oil) mixed.

Certain injection compositions are aqueous isotonic solutions or suspensions, while suppositories are advantageously prepared from fatty emulsions or suspensions. These compositions can be sterilized and/or contain adjuvants such as preservatives, stabilizers, wetting or emulsifying agents, solution enhancers, salts for adjusting osmotic pressure, and/or buffers. In addition, they may also contain other substances with therapeutic value. These compositions are prepared according to conventional mixing, granulating or coating methods, and contain about 0.1 to 75% or about 1 to 50% of active ingredients. For transdermal A suitable composition for administration contains an effective amount of a compound of the invention with a suitable carrier. Carriers suitable for transdermal delivery include pharmacologically acceptable solvents that are easily absorbed to help pass through the skin of the host. For example, the transdermal device is in the form of a bandage, which includes a support member, a reservoir containing a compound with an optional carrier, and a barrier to control the rate as needed (for a long time at a controlled and predetermined rate Delivery of host skin compounds), and methods for securing the device to the skin.

Suitable compositions for topical application (e.g., to the skin and eyes) include, for example, aqueous solutions, suspensions, ointments, creams, gels, or sprayable formulations for delivery via aerosols and the like. Such topical administration systems are particularly suitable for skin application, for example, for the treatment of skin cancer, for example, for preventive use in sunscreen creams, detergents, sprays, and the like. Therefore, they are particularly suitable for use in topical formulations known in the technical field of the invention, including cosmetics. Such can include solubilizers, stabilizers, osmolality enhancers, buffers and preservatives.

Topical administration as used herein may also involve inhalation or intranasal administration. With or without the use of suitable propellants, they can be in the form of dry powder (alone, or in a mixture, such as a dry mixture with lactose, or granules such as a mixed ingredient with phospholipids) from a dry powder inhaler or from a pressurized container, The aerosol spray of the pump, sprayer, nebulizer or atomizer is presented for convenient delivery.

Because water can promote the degradation of specific compounds, the present invention further provides anhydrous pharmaceutical compositions and dosage forms containing the compounds of the present invention as active ingredients.

The anhydrous pharmaceutical composition and dosage form of the present invention can be prepared using anhydrous or low moisture content ingredients and low moisture or low humidity conditions. The anhydrous pharmaceutical composition can be prepared and injected to maintain its anhydrous properties. Therefore, the anhydrous compositions are packaged with materials known to prevent exposure to water, so that they can be included in the appropriate package. Examples of suitable packaging materials include, but are not limited to, airtight metal foils, plastic products, unit-dose containers (such as vials), blister packs, and strip packs.

The present invention further provides pharmaceutical compositions and dosage forms comprising one or more preparations that reduce the decomposition rate of the compounds of the present invention as active ingredients. Such preparations, referred to herein as "stabilizing agents, include but are not limited to antioxidants (such as ascorbic acid), pH buffers, or salt buffers, and the like.

3. Compound

Specific examples of compounds having the general structure of formula (I) are disclosed herein.

In the first embodiment of the present invention, the compound represented by formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof, in the formula

或

； A ring is

or

；

Z is O, NR ₆ or S;

A ₁ and A ₂ are each independently CR ₁ or N;

Each case of R ₁ is hydrogen; halogen; -OH; CN; -COOC _1-6 alkyl; if necessary, C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; phenyl; amine Group; N,N -di-C _1-6 alkylamino group; optionally through halogen, phenyl, or a 5- to 6-membered heterocyclic ring with 1 to 3 heteroatoms selected from N and O (the ring is optionally (C _1-6 alkyl substituted) substituted C _1-6 alkyl; Rh; ORh; a 5- to 6-membered heteroaryl ring having 1 to 3 heteroatoms selected from N and O, which ring may optionally be amine _{Group, C 1-6} alkyl substituted by amino or hydroxy, or N -mono-C _1-6 alkylaminocarbonyl or N,N -di-C _1-6 alkylaminocarbonyl; wherein

Rh is a 5- to 6-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O and S, and the ring _{is optionally substituted with C 1-6} alkyl, -OH, or pendant oxy;

Each case of R ₂ is hydrogen; halogen; CN; -COOC _1-6 alkyl; optionally C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; amino; N,N- Di-C _1-6 alkylamino group; optionally via halogen, -OH, phenyl, or a 5- to 6-membered heterocyclic ring with 1 to 2 heteroatoms selected from N and O (the ring optionally via C _{1 -6} alkyl substitution) substituted C _1-6 alkyl; Rh; ORh; a 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, which ring may optionally be through an amino group, optionally _{Requires C 1-6} alkyl substituted by amine or hydroxy, or substituted by N -mono-C _1-6 alkylaminocarbonyl or N,N -di-C _1-6 alkylaminocarbonyl;

Each case of R ₃ is H; deuterium; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; -C(=O)NR _a R _b ; optionally halogen, -OH, C _1-6 alkyl, -NH ₂ , -NHC(=0)C _1-6 alkyl, N -di-C _1-6 alkylamino, or N -C _1-6 alkoxy substituted by mono-C _1-6 alkylamino group; optionally halogen, C _2-6 alkenyl, -OH, -NH ₂ , -NHC(=O)C _1-6 Alkyl, N -di-C _1-6 alkylamino, N -mono-C _1-6 alkylamino, -O-Rg, Rg, phenyl, or C _1-6 alkoxy substituted C _1-6 alkyl (wherein the alkoxy group is optionally substituted with halogen, -OH, C _1-6 alkoxy, N,N -di-C _1-6 alkylamino, Rg or phenyl); Requires C _3-6 ring substituted by halogen, -OH, C _1-6 alkyl, N,N -di-C _1-6 alkylamino or C _1-6 alkoxy-C _{1-6 alkyl} Alkyl; phenyl substituted by halo or C _1-6 alkoxy as required; 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, which ring may be optionally selected _{C 1-6} alkyl substituted by amine or -OH; Rg; or N,N -di-C _1-6 alkylaminocarbonyl; wherein

Rg is a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, and the ring is optionally controlled by -OH, -NH ₂ , C _1-6 alkyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution;

R _a is independently H or an optionally substituted C _1-6 alkoxy group of C _1-6 alkyl, R _b is independently H, C _1-6 alkyl, -COC _1-6 alkyl, -SO ₂ C _1-6 alkyl, C _3-6 cycloalkyl or a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, the ring is optionally connected to C _1-6 alkyl, C _{1 -6} alkoxy-C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution, or

R _a and R _b , together with the nitrogen atom to which they are connected, form a 4- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O, and S, and the ring is optionally controlled by -OH, -NH ₂ , N -Di-C _1-6 alkylamino, N -mono-C _1-6 alkylamino, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, O-cyclopropyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkyl-carbonyl substitution;

Each case of R _{c is C 1-6} alkyl or C _3-6 cycloalkyl;

Wherein the R _a, R _b, or R _c or as shown by the R _a, R _b, or the R groups depending on the alkyl group shown _c needs halogen, -OH, C _1-2 alkoxy, or C _3-4 cycloalkyl substitution;

Each case of R ₄ is H, deuterium, halogen, CN, C _1-6 alkyl, or C _1-6 haloalkyl;

Each case of R ₄ 'is H, deuterium, F, or Cl;

Each case of R ₅ is H, deuterium, C _1-6 alkyl, or C _1-6 haloalkyl; and

R ₆ is H; OH; optionally substituted with halogen, OH, C _1-6 alkoxy or C _1-6 alkyl; or an optionally halogen-substituted OH, C _1-6 alkoxy groups or C _3-6 cycloalkyl.

In the second embodiment of the present invention, the compound of formula (I) or its pharmaceutically acceptable salt or stereoisomer, wherein Z is O, and the remaining variables are as defined in the first embodiment.

，其餘變數如第一或第二具體例中所界定。 In the third embodiment of the present invention, the compound of formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the A ring is

, And the remaining variables are as defined in the first or second specific example.

In the fourth embodiment of the present invention, the compound of formula (I), or a pharmaceutically acceptable salt or stereoisomer thereof, wherein _{each example of R 3} is H; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; -C(=O)NR _a R _b ; optionally halogen, -OH, C _1-6 alkyl, -NH ₂ , -NHC(=O)C _1-6 alkyl, N -di-C _1-6 alkylamino, or N -mono-C _1-6 alkylamino substituted C _1-4 alkoxy; depending on need halogen, C _1-2 alkoxy, N - two -C _1-6 alkylamino, or N - the mono-substituted -C _1-6 alkylamino C _1-4 alkyl; C _{3- 6} cycloalkyl; or Rg, the remaining variables are as defined in the first, second and/or third specific examples.

In the fifth embodiment, the compound of formula (I) represented by structural formula (II-A) or (II-B):

或其醫藥上可接受之鹽、或立體異構物，其中R₄之各實例為H或F，其餘變數如第一、第二、第三及/或第四具體例中所界定。

Or a pharmaceutically acceptable salt or stereoisomer thereof, wherein _{each example of R 4} is H or F, and the remaining variables are as defined in the first, second, third and/or fourth specific examples.

In the sixth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein

Each case of R ₃ is H; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ;

-SO ₂ NR _c ; -C(=O)NR _a R _b ; optionally halogen, -OH, C _1-4 alkyl, -NH ₂ , N -di-C _1-4 alkylamino, or N - mono-substituted -C _1-4 alkyl of C _1-4 alkoxy group; an optionally halogen, C _1-2 alkoxy, N - two -C _1-4 alkylamino, or N -C _1-4 alkyl substituted by mono-C _1-4 alkylamino group; C _3-6 cycloalkyl group; mophorinyl group; oxetanyl group; or nitrogen heterocycle substituted by methyl group as required Butyl;

R _a is independently H or an optionally substituted C _1-4 alkoxy group, the C _1-4 alkyl;

R _{b is} independently H, C _1-4 alkyl, -COC _1-4 alkyl, -SO ₂ C _1-4 alkyl, C _3-6 cycloalkyl or having 1 to 3 selected from N and O A 3- to 6-membered heterocyclic ring with three heteroatoms, which is optionally substituted _{by C 1-4} alkyl, C _{1-4 alkoxy-C 1-4} alkyl, or C _1-4 alkoxy-carbonyl, or

R _a and R _b , together with the nitrogen atom to which they are connected, form a 4- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O, and S, and the ring is optionally controlled by -OH, C _1-4 Alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _1-4 haloalkoxy, or C _1-4 alkoxy-C _1-4 alkyl substituted, and the remaining variables are as the first , The second, the third, the fourth and/or the fifth specific example.

In the seventh embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein each example of R4 is H, _{Halogen, C 1-2} alkyl substituted by halogen as required _{, each example of R 4} is H, and the remaining variables are as in the first, second, third, fourth, fifth and/or sixth specific examples Define.

In the eighth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₂ is H, Cl, _{CN, or C 1-6} alkyl substituted by halo or -OH as needed, and the remaining variables are as specified in the first, second, third, fourth, fifth, sixth and/or seventh specific examples Define.

In the ninth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₁ is H; halogen; -OH; CN; C _1-6 alkoxy substituted by halogen as required; C _1-6 alkoxycarbonyl; phenyl; N,N -di-C _1-6 alkylamino; optionally halogen or A phenyl substituted C _1-6 alkyl group; a 5- to 6-membered heteroaryl ring containing 1 to 3 N atoms, which is optionally substituted by an amino or hydroxyl substituted C _1-6 alkyl group or by Mono- N -C _1-6 alkylaminocarbonyl or di- N -C _1-6 alkylaminocarbonyl substitution; O-Rh; or Rh; wherein Rh contains 1 to selected from N, O and S A 5- to 6-membered heterocyclic group with 4 heteroatoms, the ring may be substituted by a C _1-6 alkyl group, OH, or a pendant oxy group as necessary, and the remaining variables such as the first, second, third, fourth, fifth, Defined in the sixth, seventh, and/or eighth specific examples.

In the tenth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₁ is H; halogen; Optionally, C _1-4 alkyl substituted by halogen; C _1-4 alkoxy; or 5- to 6-membered heteroaryl containing 1 to 3 N atoms, and other variables such as first, second, third, Defined in the fourth, fifth, sixth, seventh, eighth, and/or ninth specific examples.

、

、

、

、-CH(CH₃)N(CH₃)₂、-CH₂OCH₃、環丙基、嗎福林基、-S(CH₃)、-N(CH₃)₂、-N(CH₃)(CH₂CH₃)、-N(CH₂CH₃)₂、-N(CH₃)(異丙基)、-N(CH₃)(環丙基)、-N(CH₂CH₃)(環丙基)、-N(CH₃)(SO₂CH₃)、-N(CH₂CH₂OCH₃)(環丙基)、或-N(CH₃)(CH₂CH₂OCH₃)，其餘變數如第一、第二、第三、第四、第五、第六、第七、第八、第九、及/或第十等具體例中所界定。 In the eleventh embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein each example of _{R 3 is} Bromine, methyl, ethyl, propyl, isopropyl, COOH, -CH(CH ₃ )OCH ₃ , -CH(CH ₃ )OCH ₂ CH ₃ ,

,

,

,

, -CH(CH ₃ )N(CH ₃ ) ₂ , -CH ₂ OCH ₃ , cyclopropyl, mopholinyl, -S(CH ₃ ), -N(CH ₃ ) ₂ , -N(CH ₃ ) (CH ₂ CH ₃ ), -N(CH ₂ CH ₃ ) ₂ , -N(CH ₃ )(isopropyl), -N(CH ₃ )(cyclopropyl), -N(CH ₂ CH ₃ )( Cyclopropyl), -N(CH ₃ )(SO ₂ CH ₃ ), -N(CH ₂ CH ₂ OCH ₃ )(cyclopropyl), or -N(CH ₃ )(CH ₂ CH ₂ OCH ₃ ), The remaining variables are as defined in specific examples such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, and/or tenth.

、

、乙基、或環丙基，其餘變數如第一、第二、第三、第四、第五、第六、第七、第八、第九、及/或第十等具體例中所界定。 In the twelfth embodiment, the compound of formula (I), (II-A), or (II-B) or a pharmaceutically acceptable salt or stereoisomer thereof, wherein each example of _{R 3 is -CH} (CH ₃ )OCH ₃ ,

,

, Ethyl, or cyclopropyl, other variables are as defined in specific examples such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, and/or tenth .

In the thirteenth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₄ is H, F , Cl, Br, CH ₃ , or CF ₃ , other variables such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh and / Or as defined in the twelfth class of specific cases.

In the fourteenth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₂ is Cl or CF _3. Other variables such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth and/or thirteenth Defined in the example.

In the fifteenth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₁ is H, 1 ,2,3,-Triazole, -OCH ₃ , or CF ₃ , and other variables such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, Defined in the eleventh, twelfth, thirteenth and/or fourteenth specific examples.

In the sixteenth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein R ₄ is H or Cl , Other variables such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, And/or as defined in the fifteenth and other specific examples.

為

，其餘變數如第一、第二、第三、第四、第五、第六、第七、第八、第九、第十、第十一、第十二、第十三、第十四、第十五及/或第十六等具體例中所界定。 In the seventeenth embodiment, the compound of formula (I), (II-A), or (II-B) or its pharmaceutically acceptable salt, or stereoisomer, wherein

for

, Other variables such as the first, second, third, fourth, fifth, sixth, seventh, eighth, ninth, tenth, eleventh, twelfth, thirteenth, fourteenth, Defined in the fifteenth and/or sixteenth specific examples.

In a specific example, as described herein, the compound or a pharmaceutically acceptable salt thereof is selected from compounds of formula (I), (II-A), or (II-B), or in the examples .

1. The compound represented by structural formula (I):

或其醫藥上可接受之鹽，其中

Or a pharmaceutically acceptable salt thereof, wherein

、

、

、或

； A ring is

,

,

,or

；

Z is O, N, or S;

A ₁ and A ₂ are each independently CR ₁ or N;

Each case of R ₁ is halogen; CN; -COOC _1-6 alkyl; optionally C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; amino; N,N-di- C _1-6 alkylamino; is optionally substituted by halogen, phenyl, or substituted heteroatoms selected from N and O of the 1-2 5-6 of the heterocycle C _1-6 alkyl group (the surveying requires Substituted by C _1-6 alkyl); Rh; ORh; a 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, the ring optionally via an amino group, optionally via an amino group or Hydroxy substituted C _1-6 alkyl, or substituted by N -mono-C _1-6 alkylaminocarbonyl or N,N -di-C _1-6 alkylaminocarbonyl; wherein

Rh is a 5- to 6-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O and S, and the ring _{is optionally substituted with C 1-6} alkyl, -OH, or pendant oxy,

Each case of R ₂ is halogen; CN; -COOC _1-6 alkyl; optionally C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; amino; N,N-di- C _1-6 alkylamino group; optionally halogen, phenyl, or a 5- to 6-membered heterocyclic ring with 1 to 2 heteroatoms selected from N and O (the ring is optionally substituted _{by C 1-6 alkyl} ) Substituted C _1-6 alkyl; Rh; ORh; a 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, the ring is optionally via an amino group, optionally via an amino group or hydroxy or by N - mono -C _1-6 alkylamino carbonyl or N, N - bis -C _1-6 alkyl substituted amino C _1-6 alkyl group substituted with a carbonyl group;

Where each of R ₃ is H; deuterium; _{halo; -OH; NR a R b;} -SR c; -SO 2 R c; -SO 2 NR c; is optionally substituted by halogen, -OH, C _1-6 alkoxy groups, -NH _2, N- two -C _1-6 alkylamino, or N- mono-substituted -C _1-6 alkyl group of C _1-6 alkoxy; optionally halogen, C _{2- 6} Alkenyl, -OH, -NH ₂ , N-di-C _1-6 alkylamino, N-mono-C _1-6 alkylamino, O-Rg, Rg, phenyl, or C _{1 -6} alkoxy (wherein the alkoxy group is optionally substituted by halogen, -OH, C _1-6 alkoxy, N,N-di-C _1-6 alkylamino, Rg or phenyl)的C _1-6 alkyl; optionally halogen, -OH, C _1-6 alkyl, N,N -di-C _1-6 alkylamino or C _1-6 alkoxy-C _{1-6 C 3-6} cycloalkyl substituted with alkyl; phenyl substituted with halo or C _1-6 alkoxy if necessary; 5 to 6 membered heteroaromatic with 1 to 3 heteroatoms selected from N and O Rg; or N,N-di-C _1-6 alkylaminocarbonyl group; wherein the ring is _{optionally substituted with a C 1-6} alkyl group substituted by an amine group or -OH as needed; Rg;

R _g is a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, and the ring is optionally controlled by -OH, -NH ₂ , C _1-6 alkyl, C _1-6 alkoxy- C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution;

R _{a is} independently H or C _1-6 alkyl, and R _{b is} independently H, C _1-6 alkyl, -COC _1-6 alkyl, -SO ₂ C _1-6 alkyl, C _{3- A 6-} cycloalkyl group or a 3- to 6-membered heterocyclic ring with 1 to 3 heteroatoms selected from N and O, the ring is optionally controlled _{by C 1-6} alkyl, C _{1-6 alkoxy-C 1-6} alkane Group, or C _1-6 alkoxy-carbonyl substitution), or

R _a and R _b , together with the nitrogen atom to which they are connected, form a 4- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O, and S, and the ring is optionally controlled by -OH, -NH ₂ , N-di-C _1-6 alkylamino, N-mono-C _1-6 alkylamino, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _1-6 haloalkoxy, O-cyclopropyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkyl-carbonyl substitution;

Each case of R _{c is C 1-6} alkyl or C _3-6 cycloalkyl;

Wherein the R _a, R _b, or R _c or as shown by the R _a, R _b, or the R groups depending on the alkyl group shown _c needs halogen, -OH, C _1-2 alkoxy, or C _3-4 cycloalkyl substitution;

Each case of R ₄ is H, deuterium, halogen, CN, C _1-6 alkyl, or C _1-6 haloalkyl; and

Each case of R ₅ is H, deuterium, C _1-6 alkyl, or C _1-6 haloalkyl.

2. The compound of paragraph 1 or a pharmaceutically acceptable salt thereof, wherein _{each example of R 3} is H; halo; -OH; NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; C substituted by halogen, -OH, C _1-6 alkyl, -NH ₂ , N-di-C _1-6 alkylamino, or N-mono-C _1-6 alkylamino as needed _1--4 alkoxy; optionally substituted halo C _1-2 alkoxy groups or C _1-4 alkyl; C _3-6 cycloalkyl; or Rg.

3. The compound of paragraph 1 or 2, or a pharmaceutically acceptable salt thereof, wherein Z is O.

4. The compound of any one of paragraphs 1 to 3, or a pharmaceutically acceptable salt thereof, wherein R ₃ is H; halo; -OH; NR _a R _b ; -SR _c ; -SO ₂ R _c ;- SO ₂ NR _c ; optionally halogen, -OH, C _1-6 alkyl, -NH ₂ , N-di-C _1-6 alkylamino, or N-mono-C _1-6 alkylamino Substituted C _1-4 alkoxy; methyl, ethyl, isopropyl, C _1-4 haloalkyl; morpholinyl; oxetanyl; or, if necessary, azetidine substituted by methyl base.

。 5. The compound of any one of paragraphs 1 to 4, or a pharmaceutically acceptable salt thereof, wherein the A ring is

.

6. The compound of any one of paragraphs 1 to 5, wherein the compound is represented by the structural formula (II-A) or (II-B):

或其醫藥上可接受之鹽。

Or its pharmaceutically acceptable salt.

7. The compound of any one of paragraphs 1 to 6, or a pharmaceutically acceptable salt thereof, wherein _{each case of R 4} is H, halogen, and optionally C _1-2 alkyl substituted with halo.

8. The compound of any one of paragraphs 1 to 7, or a pharmaceutically acceptable salt thereof, wherein R ₂ is H, Cl, CN, or C _1-6 alkyl substituted with halo or -OH as necessary.

9. The compound of any one of paragraphs 1 to 8, or a pharmaceutically acceptable salt thereof, wherein R ₁ is H; halogen; -OH; CN; C _1-6 alkoxy substituted by halogen as necessary;

C _1-6 alkoxycarbonyl group; phenyl group; N,N-di-C _{1-6 alkylamino group; C 1-6} alkyl group substituted by halogen or phenyl group as required; containing 1 to 3 N atoms A 5- to 6-membered heteroaryl ring, which is optionally substituted by a C _1-6 alkyl group substituted by an amino group or a hydroxy group, or substituted by a mono- or di- N -C _1-6 alkylaminocarbonyl group as required ; O-Rh; or Rh; wherein Rh is a 5- to 6-membered heterocyclic group containing 1 to 4 heteroatoms selected from N, O and S, and the ring is optionally connected by C _1-6 alkyl, OH, or side Oxy-substituted.

10. The compound of any one of paragraphs 1 to 9, or a pharmaceutically acceptable salt thereof, wherein R ₁ is H; halogen; optionally C _1-4 alkyl substituted by halogen; or containing 1 to 3 N Atomic 5 to 6 membered heteroaryl group.

11. The compound of any one of paragraphs 1 to 10, or a pharmaceutically acceptable salt thereof, wherein _{each case of R 3} is H, ethyl, isopropyl, cyclopropyl, morpholinyl, N(CH ₃ ) (Cyclopropyl) or (CH ₃ )(CH ₂ CH ₂ OCH ₃ ).

12. The compound of any one of paragraphs 1 to 10, or a pharmaceutically acceptable salt thereof, wherein _{each case of R 3} is H, F, or Cl, preferably H or F.

13. The compound of any one of paragraphs 1 to 12, or a pharmaceutically acceptable salt thereof, wherein R ₄ is H, F, Cl, Br, CH ₃ , or CF ₃ .

14. The compound of any one of paragraphs 1 to 13, or a pharmaceutically acceptable salt thereof, wherein R ₂ is Cl or CF ₃ .

15. The compound of any one of paragraphs 1 to 14, or a pharmaceutically acceptable salt thereof, wherein R ₁ is H, 1,2,3,-triazole, or CF ₃ .

16. The compound of any of paragraphs 1 to 15, or a pharmaceutically acceptable salt thereof, wherein R ₄ is H or Cl.

為

。 17. The compound of any of paragraphs 1 to 16, or a pharmaceutically acceptable salt thereof, wherein

for

.

In a specific embodiment, the compound of the present invention is a partial inhibitor of MALT1, in which a maximum of about 50 to 85%, or a maximum of about 60 to 80%, or a maximum of about 70 to 80%, or a maximum of about 75% of MALT1 activity is inhibited . Inhibition of MALT1 activity can be evaluated using any method recognized in the technical field of the invention, for example, in the biological examples, especially as described in Example 1 (Glosensor assay or its variants) and/or Example 2 (IL-2 production) By. In a specific example, the Glosensor assay as described in Biological Example 1 was used to evaluate MALT1 activity.

4. Treatable diseases

In specific embodiments, the compounds of the present invention can be used to treat the activity of the CBM complex (e.g., one of CARMA1/BCL10/MALT1 or CARD9/BCL10/MALT1) and/or MALT1 activity is up-regulated or overactive (e.g. , Activating CARD11 mutation, wild-type MALT1 overexpression, activating mutation of MALT1 function, loss of function of MALT1 inhibitor, etc.) diseases or indications.

Thus in particular embodiment, the compounds of the present invention may be used to treat NF- κ B Messaging wherein the disease or disorder or indication of the path is activated. While not wishing to be bound by any particular theory of, but is generally believed to such a disease or condition (e.g., cancer), the direct blocking or inhibiting NF- κ B MALT1 adjusted downward path, thereby leading to treatment.

In another particular embodiment, although still not wishing to be bound by any particular theory of, but is further believed that, the MALT1 inhibitors of the present invention may not be in certain tumor cells, e.g. deregulated NF- κ B inhibition of the direct path and dominated kick in. Conversely, inhibition effect on caspase activity bit relating MALT1 independent or compound comprising a plurality of path components NF- κ B immune systems, and a wide regulation of specific T cell populations subject, e.g. Th17 cells, or Natural regulatory T cells (nTreg) or Tregs, but not the other, or both. Therefore, certain compounds of the present invention act as immunomodulators to fine-tune the activity of specific T cell populations, and indirectly expand the range of treatments for cancer and other autoimmune/inflammatory diseases or indications by administering MALT1 inhibitors to subjects , regardless of such cancer / autoimmune / inflammatory disease or disorder indication of whether NF- κ B activity path.

In specific examples, the compounds of the present invention preferentially or selectively inhibit Th17 cells relative to Treg (for example, relative to Th17, the IC 50 ratio of Treg exceeds 3, 5, 10, 15, 20, 25 _{, 30, 40, 50} , 60, 70, 80, or 100). In certain specific examples, the compounds of the present invention do not substantially inhibit Treg.

The symptoms and diseases characterized by NF-kB activation disorder include autoimmune, immune, and/or inflammatory diseases, allergic diseases, respiratory diseases, and tumor diseases.

In one aspect, the present invention provides a method for inhibiting MALT1 activity in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a compound of the present invention.

In another related aspect, the compounds of the present invention can be used to manufacture agents that inhibit the activity of MALT1.

According to any of the above related aspects of the present invention, the subject has a disease or symptom, such as an autoimmune disease, inflammatory disease, or cancer, and wherein the disease or symptom is treated.

In specific cases, the disease or symptoms are rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE), vasculitis, allergic diseases, respiratory diseases [e.g. Asthma and chronic obstructive pulmonary disease (COPD)], symptoms caused by delayed or immediate allergic reactions, acute allergies, acute or chronic transplant rejection, graft-versus-host disease, cancers of hematopoietic origin, or solid tumors, including chronic types Myelogenous leukemia (CML), myeloid leukemia, non-Hodgkin’s lymphoma (HL), or B-cell lymphoma.

In a specific example, the treatable disease or condition is cancer (tumor disease).

Tumor diseases that can be treated with the compounds of the present invention can include, in particular, cancers of hematopoietic origin or solid tumors, malignant tumors, sarcomas, lymphomas, leukemias and germ cell tumors (e.g. adenocarcinoma), bladder cancer, clear cell carcinoma, skin cancer, brain Cancer, cervical cancer, colon cancer, colorectal cancer, endometrial cancer, bladder cancer, brain tumor, breast cancer, gastric cancer, germ cell tumor, glioblastoma, liver adenoma, Hodgkin's Lymphoma, liver cancer, kidney cancer, lung cancer, ovarian cancer, dermal tumor, prostate cancer, renal cell carcinoma, stomach cancer, neuroblastoma, non-Hodgkin's lymphoma, diffuse large B cell Lymphoma, adventitia cell lymphoma, border zone lymphoma, skin T-cell lymphoma, other B-cell lymphoma, melanoma, gastric mucosa-associated lymphoma (MALT) lymphoma, chronic myelogen-like leukemia, myeloid Leukemia, multiple myeloma, plasma cell tumors, mollusc melanoma, and acral mollusc melanoma.

In certain specific cases, the cancer is leukemia or lymphoma.

In a specific example, the leukemia is chronic lymphocytic leukemia (CLL), such as CLL with CARD11 mutation.

In certain specific cases, the lymphoma is gastric mucosa-associated lymphoid tissue lymphoma (MALT).

In certain specific cases, the lymphoma is adventitia cell lymphoma. In certain specific cases, the lymphoma is a border zone lymphoma. In certain specific cases, the lymphoma is cutaneous T-cell lymphoma such as Sezary syndrome. In certain specific cases, the lymphoma is primary exudate lymphoma.

In certain specific cases, the lymphoma is non-Hodgkin's lymphoma (NHL). In certain specific cases, the lymphoma is DLBCL (diffuse large B-cell lymphoma). In certain specific cases, the lymphoma is germinal center B cell (GCB) DLBCL. In a specific example, the lymphoma is activated B cell (ABC) DLBCL (for example, a person with an activating mutation in CARD11).

In certain specific cases, the cancer is lung adenocarcinoma.

In certain specific cases, the cancer is breast cancer.

In certain specific cases, the cancer is pancreatic cancer.

The autoimmune and/or inflammatory diseases that can be treated with the compounds of the present invention can be especially selected from arthritis, ankylosing spondylitis (AS), inflammatory bowel disease (IBD), ulcerative colitis (UC), gastritis , Pancreatitis, Crohn's disease (CD), diarrheal disease, multiple sclerosis (MS), systemic lupus erythematosus (SLE), rheumatic fever, gout, organ or transplant rejection, acute or chronic graft Vs. host disease, chronic allograft rejection, Behcet's disease, uveitis, psoriasis, dermatitis, atopic dermatitis, dermatomyositis, myasthenia gravis, Graves' disease ( Grave's disease, Hashimoto's thyroiditis, Shogren's syndrome, and vesicular disorders (such as pemphigus vulgaris), antibody-mediated vascular inflammation syndrome (including ANCA-related vasculitis, Hennoch-Schonlein) Purple scar disease and immunity Complex vasculitis (regardless of primary or secondary infection or cancer)].

In certain specific cases, the treatable disease or disorder is an autoimmune disease or disorder, or an inflammatory disease or disorder.

In certain specific cases, the disease is rheumatoid arthritis (RA).

In certain specific cases, the disease is psoriasis.

In certain specific cases, the disease is ulcerative colitis (UC).

In certain specific cases, the disease is multiple sclerosis (MS).

In certain specific cases, the disease is allergic encephalomyelitis.

In certain specific cases, the disease is systemic lupus, Sogren’s syndrome, or Hashimoto’s thyroiditis.

In certain specific cases, the disease or condition is an allergy caused by chronic inflammation.

Allergies that can be treated with the compounds of the present invention include contact dermatitis, diarrhea diseases, asthma, allergies to house dust mites, pollen and related allergens, beryllium poisoning [or chronic beryllium disease (CBD)].

Respiratory system diseases that can be treated with the compounds of the present invention, especially including: asthma, bronchitis, chronic obstructive pulmonary disease (COPD), cystic fibrosis, pulmonary edema, pulmonary embolism, pneumonia, pulmonary sarcoidosis, silicosis Disease, pulmonary fibrosis, respiratory failure, acute respiratory distress syndrome, primary pulmonary circulatory hypertension and emphysema.

In other specific examples, the compounds of the present invention can also be used to treat BENTA ("B cell expansion with NF-κB and T cell anergy") diseases, lupus nephritis, and polymyositis.

In certain specific cases, the autoimmune disease or disease, or the inflammatory disease or disease is further administered by immunosuppressive agents, such as cyclosporine, rapamycin, Methotrexate and other treatments.

5. Combination therapy

The compounds of the present invention can be used in combination therapy with one or more additional/adjuvant therapeutic agents suitable for treating diseases or indications that the subject compounds can treat.

Therefore, in a specific embodiment, the method of the present invention using the compound of the present invention may additionally include administering a further therapeutic agent to a subject in need thereof. The further therapeutic agent may be: (i) an immunomodulator that blocks or inhibits immune system checkpoints, which may or may not be a component of the NF-κB pathway; and/or (ii) directly stimulate immune effectors Reactive agents, such as cytokines, or tumor-specific adoptively transferred T cell populations, or antibodies specific to proteins expressed by tumor cells; and/or (iii) compositions containing tumor antigens or immunogenic fragments thereof ; And/or (iv) chemotherapeutic agents.

The compounds of the invention can be administered simultaneously, before or after the further therapeutic agent. MALT1 inhibitors can be administered separately via the same or different routes of administration, or used together as further therapeutic agents in the same pharmaceutical composition.

"Co-administration" or "co-administration" as used herein is intended to cover the administration of the selected therapeutic agent to a single patient, and is intended to include a treatment plan in which the preparations are not necessarily administered via the same route of administration or at the same time.

The term "pharmaceutical composition" as used herein means a product produced by mixing or combining more than one active ingredient, and includes both fixed and non-fixed combinations of the active ingredients. The term "fixed composition" means that the active ingredients (for example, the subject compound and a co-formulation) are simultaneously administered to the patient in the form of a single unit or dosage. The term "non-fixed composition" means that the active ingredients (for example, the subject compound and the co-formulation) are both in separate entities, without a specific time limit, and are administered to the patient simultaneously, concurrently or sequentially, wherein such administration provides these The therapeutically effective amount of the compound in the patient's body.

In one specific example, the present invention provides a product containing the compound of the present invention (for example, the subject compound or any subgroup thereof), and at least one other therapeutic agent as a combined preparation for simultaneous, separate or sequential use in treatment. The product provided as a combination preparation includes a composition comprising the compound of the present invention or any subgroup thereof and other therapeutic agents together in the same pharmaceutical composition, or the subject compound or the subject compound in a separate form (for example, in the form of a kit) Any subgroup and other therapeutic agents.

In a specific example, the present invention provides a medical composition for use in therapy, which comprises a subject compound or any subgroup thereof and another immunomodulator or a composition comprising a tumor antigen or an immunogen fragment thereof. Optionally, the pharmaceutical composition may contain pharmaceutically acceptable excipients.

In a specific example, the present invention provides a kit comprising two or more different pharmaceutical compositions, at least one of which contains the subject compound, and the other contains the auxiliary therapeutic agent discussed herein. In a specific example, the kit includes a means for storing the composition separately, such as a container, a separate bottle, or a separate metal foil packet. An example of this type of kit is blister packaging, which is commonly used as packaging material for tablets, capsules and the like. The kit of the present invention can be used to administer different dosage forms (for example, oral and parenteral), to administer separate compositions at different dosage intervals, or to titrate the separate compositions with each other. To assist compliance, the kits of the present invention usually include dosing guidelines.

It should be understood that many of the further therapeutic agents used in the method of the present invention may be biological products that require intravenous, intraperitoneal or injection administration (depot administratio n). In further specific examples, the compound of the present invention is administered orally, and the further therapeutic agent is administered parenterally (for example, intravenously, intraperitoneally, or as an injection).

In certain specific cases, the second/additional therapeutic agent is an immune system checkpoint. The activation of effector T cells is usually triggered by the TCR recognition antigenic peptide presented by the MHC complex. Then, through the balance between the stimulus signal and the inhibitory effector T cell response signal, the type and degree of activation achieved are determined. As used herein, "immune system checkpoint" refers to any molecular interaction that alters the balance in favor of suppressing the response of effector T cells. That is, when molecular interaction occurs, it negatively regulates the activation of effector T cells. Such interactions may be direct, such as interactions between ligands and cell surface receptors that transmit inhibitory signals to effector T cells. Or it may be indirect, such as blocking or inhibiting the interaction between the ligand and the cell surface receptor, otherwise the receptor will send activation signals to effector T cells, or promote the interaction of inhibitory molecules or cell upregulation. Or the metabolites required by effector T cells are reduced by enzymes, or any combination thereof.

Examples of immune system checkpoints include: a) the interaction between indoleamine 2,3-dioxygenase (ID01) and its substrate; b) the interaction between PD1 and PD-L1 and/or PD1 and PD-L2; c) Interaction between CTLA-4 and CD86 and/or CTLA-4 and CD80; d) Interaction between B7-H3 and/or B7-H4 and their respective ligands; e) Interaction between HVEM and BTLA; f) Interaction between GAL9 and TIM3; g) Interaction between MHC Type I or Type II and LAG 3; and h) Interaction between MHC Type I or Type II and KIR; i) OX40( The interaction between CD134) and OX40L (CD252); j) the interaction between CD40 and CD40L (CD154); k) the interaction between 4-1 BB (CD137) and ligands (including 4-1 BBL); 1 ) Interaction between GITR and ligands (including GITRL).

The representative checkpoint used for the purpose of the present invention is checkpoint (b), which is the interaction between PD1 and any of its ligands PD-L1 and PD-L2. PD1 is expressed on effector T cells. Its binding to any ligand will generate a signal that activates down-regulation. Ligands are manifested by certain tumors. PD-L1 is particularly manifested by many solid tumors (including melanoma). Therefore, these tumors may down-regulate the anti-tumor effect of the immune vector by activating the inhibitory PD1 receptor on T cells. By blocking the interaction between PD1 and one or two of its ligands, the checkpoint of the immune response can be removed, resulting in an enhanced anti-tumor T cell response. Therefore, PD1 and its ligands can be the actual components of the immune system checkpoint targeted in the method of the present invention. example.

Another checkpoint used for the purpose of the present invention is checkpoint (c), which is the interaction between the T cell receptor CTLA-4 and its ligand B7 protein (B7-1 and B7-2). After initial activation, CTLA-4 is usually upregulated on the surface of T cells and binds to the ligand to generate a signal that inhibits further/sustained activation. CTLA-4 competes with the receptor CD28 for binding to the B7 protein. The receptor CD28 also appears on the surface of T cells but up-regulates activation. Therefore, by blocking the interaction between CTLA-4 and B7 protein instead of the interaction between CD28 and B7 protein, one of the normal immune response checkpoints can be removed, resulting in an enhanced anti-tumor T cell response. Therefore, CTLA-4 and its ligand can be an example of the target immune system checkpoint component in the method of the present invention.

The "immunomodulator" as used herein includes any preparation that blocks or inhibits the checkpoint of the immune system when administered to a subject, leading to an upward regulation of the subject's immune effector response. The immune effector response is usually T Cell effector responses, which may include anti-tumor T cell effector responses.

The immunomodulators used in the method of the present invention can block or inhibit any of the above-mentioned immune system checkpoints. The preparation may be an antibody or any other suitable preparation that causes the blocking or inhibition. Therefore, the preparation is often referred to as an inhibitor of the checkpoint.

"Antibody" as used herein includes whole antibodies and any antigen-binding fragments (ie, "antigen-binding portions") or single chains thereof. The antibody can be a multi-strain antibody or a monoclonal antibody, and can be produced via any appropriate method. Examples of binding fragments covered by the term "antigen-binding portion" of antibodies include Fab fragments, F(ab') ₂ fragments, Fab' fragments, F _d fragments, Fv fragments, dAb fragments, and isolated complementarity determining regions (CDR). Single-chain antibodies (such as scFv) and heavy-chain antibodies (such as VHH and camelid antibodies) are also intended to be encompassed by the term "antigen-binding portion" of antibodies.

In certain specific cases, the anti-inflammatory agent that blocks or inhibits the interaction of CTLA-4 and B7 protein The body includes Ipilimumab, tremelimumab, or any antibody disclosed in WO2014/207063. Other molecules include polypeptides, or soluble mutant CD86 polypeptides. In a specific embodiment, the antibody is ipilimumab.

In specific cases, antibodies that block or inhibit the interaction of PD1 and PD-L1 include Nivolumab, Pembrolizumab, Lambrolizumab, and Pitilizumab. Anti (Pidilizumab), BGB-A317 and AMP-224. In certain embodiments, the antibody is nivolumab or pembrolizumab. Anti-PD-L1 antibodies include atezolizumab, avelumab or durvalumab, MEDI-4736 and MPDL3280A.

In certain embodiments, antibodies that block or inhibit the interaction between 4-1BB and its ligand include utomilumab.

Other suitable inhibitors include small molecule inhibitors (SMI), which are usually small organic molecules. In certain specific examples, ID01 inhibitors include Epacadostat (INCB24360), Indoximod, GDC-0919 (NLG919) and F001287. Other inhibitors of ID01 include 1-methyltryptophan (1-MT).

As used herein, "agent that directly stimulates immune effector response" means any appropriate agent, but generally refers to cytokines or chemokines (or agents that stimulate any production), tumor-specific adoptively transferred T cell populations, Or antibodies specific to the protein expressed by tumor cells.

The cytokines may be interferons or interleukins (such as IL-2) selected from the group consisting of IFN α , IFN β , IFN γ and IFNA. Chemokines can be inflammatory mediators, such as inflammatory mediators selected from CXCL9, 10, and 11, which attract T cells that express CXCR3. Agents that stimulate the production of cytokines or chemokines may be adjuvants suitable for human administration. One example is Bacille Calmette-Guerin (BCG), which is usually administered in the bladder (ie, catheter) for the treatment of bladder cancer. The typical dosing regimen of BCG for bladder cancer is once a week for six weeks, but its long-term safety history is known, and it can also be administered indefinitely for maintenance. It has been proven that BCG stimulates the immune response to bladder cancer. BCG has also been used as an adjuvant in combination with a tumor antigen-containing composition (ie, with a cancer vaccine), especially for colon cancer, when it was usually administered intracutaneously. Such use of BCG is also planned in the present invention. The tumor-specific adoptively transferred T cell population directly increases the size of the tumor-specific T cell population in an individual, and can be produced by any appropriate means. However, usually the process involves isolating tumor-specific T cells from a tumor sample taken from a patient, and selectively culturing these cells before returning the expanded tumor-specific T cell population to the patient . Alternatively, genetic engineering of the T cell receptor locus can be followed by expansion of the altered cells to produce tumor-specific T cell populations.

Antibodies specific to proteins expressed by tumor cells usually promote cell destruction through binding to tumor cells and through antibody-dependent cellular cytotoxicity (ADCC) to stimulate immune activity. Examples of antibodies of this type include anti-CD20 antibodies [e.g. ofatumumab or rituximab] and anti-CD152 antibodies [e.g. alemtuzumab].

Therefore, in specific illustrative examples, the compounds of the present invention can be combined with calcineurin inhibitors (such as cyclosporine A or FK 506), mTOR inhibitors (such as rapamycin, 40-0-(2-hydroxyethyl) (Base) rapamycin, biolimus-7 or biolimus-9), immunosuppressive ascomycin (e.g. ABT-281, ASM981), corticosteroids, cyclophosphamide, Azathioprine, methotrexate, Ayanine, mizoribine, mycophenolic acid or salt, mycophenolate mofetil, and IL-1 β inhibitor are used in combination.

In another embodiment, the compound of the present invention is combined with a co-formulation that is a PI3 kinase inhibitor.

In another specific example, the compound of the present invention has an effect on BTK [Bruton’s (Bruton's) Tyrosine Kinase] co-preparation combination.

In order to treat tumor diseases, the compounds of the present invention can be used in combination with B cell modulators, for example, rituximab, BTK or Syk inhibitors, PKC, PI3 kinase, PDK, PIM, JAK and mTOR inhibitors and BH3 mimics .

6. Compound screening / determination methods

The compounds of the present invention inhibit the para-apoptotic protease activity of MALT1, which can be directly measured using many biochemical assays, such as the GLOSENSOR ^TM assay described in Example 1. Thus, IC ₅₀ values may be of any of the compounds at a range of inhibitor concentrations to be measured.

In Example 1 (the entire contents of which are incorporated herein, but only briefly described here to avoid redundancy), the GLOSENSOR ^TM (Promega Corp., Madison, WI) segmented luciferase system can be used to evaluate MALT1 inhibition The activity of the agent. In this system, as described in Fontan et al. ( J Clin Invest. 128(10): 4397-4412, 2018, incorporated herein for reference), the firefly luciferase is genetically modified and split into short Two different domains connected by the MALT1 matrix peptide (for example, the MALT1 matrix-the cleavage site sequence of RelB). The luciferase is inactive in this configuration; after activation of MALT1, for example, using PMA/IO (ionomycin) to stimulate cells with functional MALT1, the MALT1 matrix peptide is cleaved, resulting in a conformational change, thereby restoring A functional luciferase protein. When an appropriate luciferase substrate is provided, such as Promega Corp's Bright Glo substrate, the luminescence signal can be detected. The presence of the MALT1 inhibitor in this system reduces the luminescence signal in a manner proportional to the concentration of the MALT1 inhibitor. _{Therefore, the IC 50} value of the inhibitor can be determined at a series of inhibitor concentrations. This assay can be performed in any suitable cell line, such as Raji lymphoma cells. ^{The GLOSENSOR TM} construct encoding the luciferase enzyme can be stably integrated into the genome of the cell line.

Example 2 (the entire content of which is incorporated herein, but only briefly described here to avoid redundancy) describes alternative or more assays for evaluating the inhibition of MALT1 protease by subject compounds. According to this assay system via such inhibitors leads to production of IL-2 (for the activation of downstream events MALT1) ability to antagonize the activation of B MALT1 Communications of NF- κ. In short, T cell TRANSACT ^TM reagent (Miltenyi Biotec) (this reagent has been developed as a ready-to-use reagent to activate and expand human T cells through co-stimulation of CD3 and CD28) can be used to stimulate the production of IL- 2 appropriate cell lines [e.g. Jurkat cells (immortal human T lymphocytes)] to produce IL-2. The IL-2 thus produced can be collected from the cell supernatant, and then the amount of IL-2 can be determined based on a standard curve generated using a series of known concentrations of IL-2 using a standard ELISA assay. In this system, the presence of a MALT1 inhibitor reduces the amount of IL-2 produced in a manner proportional to the concentration of the MALT1 inhibitor. _{Therefore, the IC 50} value of the inhibitor can be determined under a series of inhibitor concentrations.

IC ₅₀ values can be determined using the subject compounds, one or both assays.

More assays can also be used to evaluate the ability of any specific MALT1 inhibitor of the present invention to inhibit the para-apoptotic protease activity of MALT1, or to screen for compounds possessing MALT1 inhibitory activity.

For example, in an assay, a fluorescent probe covalently attached to a short MALT1 matrix peptide can be used to measure the MALT1 proteolytic activity. The probe is non-fluorescent in this state, but after the matrix peptide is cleaved by activated MALT1, the probe is released and becomes fluorescent, which can be measured with a fluorometer. The presence of MALT1 inhibitors will reduce this type of fluorescence signal at a range of inhibitor concentrations.

In particular, the C domain of MALT1 (amino acids 329 to 824) can be used as a substitute for MALT1 in this inhibitor screening. The read-out parameter can be the increase in fluorescent life over time in proportion to the enzyme activity. This assay uses a short peptide matrix of MALT1, and uses a single fluorophore PT14 label as a fluorescent lifetime probe that is sensitive to the cleavage state of the matrix [PT14: 6-(9-oxo-9H-acridin-10-yl )-Caproic acid ester, AssayMetrics, UK). The peptide matrix has the following sequence: Ac-Trp-Leu-Arg-Ser-Arg-Cys(PT14)-NH ₂ (product number BS-91 17, Biosynta, Germany). Here, Ac represents an acetyl group, Cys(PT14) is a cysteine residue, and the fluorophore PT14 is joined to the sulfhydryl group of cysteine via the maleimide group; the C of the peptide The end is aminated; the cleavable bond is between Arg and Cys at the end. The buffer for the assay is composed of 200mM Tris/HCI, pH 7.5, 0.8M sodium citrate, 100 μM EGTA, 100 μM DTT, and 0.05% (w/v) CHAPS. The analysis of the kinetic characteristics of the enzyme reaction can be used to determine the Michaelis constant (K _M ) and k _cat value. It can be measured in a 384-well culture plate format using a black microtiter round-bottomed culture plate (product number 95040020, Thermo Electron Oy, Finland). The test compound can be dissolved in 100% (v/v) DMSO or a mixture containing 90% (v/v) DMSO and 10% (v/v) water, and the concentration of the stock solution is 100 mM. 100% (v/v) DMSO or a mixture containing 90% (v/v) DMSO and 10% (v/v) water can be used for serial dilution of test compounds.

To measure the inhibitory effects of compounds at the 384-well plates in the 0.25 μ L L of the test compound mixed with enzyme 12.5 μ, the mixture was incubated at room temperature (22 ℃) 60 min. Then, 12.5L of substrate was added, and the enzyme reaction was allowed to continue for 60 minutes at room temperature (22°C). Thus a total assay volume of 25.25 μ L, final assay concentration of substrate and enzyme were 2.5nM and 1 μ M. Under this measurement condition, the increase in the measurement signal over time is expected to be linear for at least 60 minutes and is proportional to the active enzyme concentration of at least 2.5 nM. The DMSO content is between 0.9 and 1% (v/v). Diluted in serial dilution factor of 3.16 in the final assay concentration of test compound generally in the range to 100 μ M of 1nM (i.e. half log dilution procedure). As a control, an uninhibited enzyme reaction (ie 0% inhibition) will be caused by adding only DMSO instead of the test compound, or by adding an assay buffer that does not contain enzymes mixed with DMSO, which is equivalent to a completely inhibited reaction (ie 100%). % Inhibition), the reaction is carried out in multiple wells. Use a microtiter plate reader, such as a TECAN Ultra Evolution FLT instrument with excitation fluorescence at 405 nm and emission recording at 450 nm, to record the fluorescence lifetime. Using the above-mentioned control as a reference (0 and 100% inhibition), the fluorescence lifetime can be converted into a percentage of inhibition. Using non-linear regression analysis software (Origin, OriginLab Corpo than n, USA), IC ₅₀ value _was calculated from the percentage of inhibition of FIG inhibitor concentration. Fit the data using a four-parameter logarithmic model, which is characterized by the following equation:

y=A2+(A1-A2)/(1+(x/IC ₅₀ )^p) where y is the% inhibition at the inhibitor concentration, A1 is the minimum inhibition value, and A2 is the maximum inhibition value. The index p is the Hill coefficient.

In another specific example, a reporter gene (such as luciferase) can also be used to evaluate MALT1 inhibition under the control of a promoter that responds to NF-kB activation. For example, the construct encoding the cIAP2-MALT1 fusion (persistent active MALT1 protease) can be firmly integrated into the host cell (for example, HEK293) gene body, and can be included in the promoter of the NF-kB response element Under control, use luciferase to read the activated NF-kB signal. As reflected by the reduced activity of the luciferase reporter gene, the presence of MALT1 inhibitors will down-regulate the NF-kB signaling stimulated by MALT1.

More specifically, this drive cIAP2-MALT1 of NF- κ B reporter gene assay (the RGA) in an appropriate cell line, for example, HEK293 performed. In this assay, the fusion protein cIAP2-MALT1 drives persistent NF-kB activation in MALT-type B-cell lymphoma. In order to monitor the activity of MALT1 inhibitors on NF-kB signaling, a stably transfected HEK293 cell line was established, in which the activated cIAP2-MALT1 fusion protein was continuously expressed, and the firefly luciferase reporter gene Under the control of NF-kB reaction element. In this system, inhibition of MALT1 protease activity reduces NF-kB signaling, and luciferase activity is reported to be low. The luciferase activity detection assay can be used to measure the inhibition of luciferase gene expression. Briefly, inoculated with 1.8 × 10 ⁴ cells / 90 μ L / well culture treated 96 well microtiter plate of (Costar, Catalog No. 3903) at a sterile, white walls, clear bottom of the organized. After 37 ° C incubated overnight using a liquid handling robot (Velocity Bravo 11, Agilent), initially prepared in DMSO followed by 1: L Add 10 μ 100 intermediate was diluted in culture medium the cells were 10 × 3-fold serial compound dilutions To the cell. Unless mentioned otherwise, the starting concentration of the compound may be set at 10 μ M, and the final concentration in all wells of the vehicle may be 0.1% DMSO. After 24 hours of incubation compounds, a first step is added to 10 μ L in phosphate buffered saline of 135 μ gmL resazurin sodium salt (SIGMA Cat-Nr R7017) 3 hours to assess cell viability. After reading the count microplate multi-use (e.g. Infinite M200Pro, TECAN), in the 540 590nm excitation / / emission wavelength quantitative reduction of resazurin, a cell with 70 μ L ONEGlow (Promega, catalog number E6120) homogeneous The assay buffer was incubated together at room temperature for 20 minutes, and then the luciferase expression level was quantified. The luminescence is recorded in the luminescence detection mode on a multi-purpose micro-disk reader (for example, Infinite M200Pro, TECAN). Use the Excel analysis template to process the raw data. The effect of a specific test compound concentration on NF-kB activity is expressed as luciferase signal (relative light unit), which is divided by the reduced resazurin signal (relative fluorescence unit) to normalize to cell viability. The value obtained from the cells treated with the vehicle is set to 100%. Using a four-parameter curve fit (XLfit, V4.3.2) determines the absolute (relative to the vehicle control group by 50%) and the relative (turning point) IC50 value (μ M). In addition, the% normalized NF-κB signal and% cell viability at the highest compound concentration were tested.

In another specific example, the human IL-2 promoter reporter gene analysis (RGA) can also be used to evaluate MALT1 inhibition in Jurkat cells. Similar to the assay in Example 2, Jurkat cells can produce IL-2 through stimulation with anti-CD28 mAb and PMA. Different from Example 2, under the control of an IL-2 response promoter, a reporter gene (such as luciferase) is used to indirectly measure the produced IL-2. Therefore, the amount of IL-2 produced is proportional to the measured luciferase activity.

In this assay, the transfected Jurkat clones can be propagated in RPMI 1640 medium supplemented with 10% heat-inactivated fetal bovine serum, 50 μM 2-mercaptoethanol and 1 mg/ml geneticin. During the culture period, the cell concentration should not exceed 1×10 ⁶ cells/ml. Cells should not be subcultured more than 30 times. Before the measurement, the cells were washed and prepared to a concentration of 2×10 ⁶ cells/mL. The compound dilution was made into a 2× concentrated solution, and then diluted 1/2 by adding to the cells. In 96 deep well culture plate, mixed with about 250 μ L of compound dilutions and 250 μ L cells. Incubate the cell/compound premix directly in a deep-well culture dish at 37°C and 5% CO _{2 for 30 minutes.} Cells after preincubation with the compound to 3g / mL of anti-CD28 mAb + 1 μ g / mL of PMA-stimulated cells. The two costimulators were diluted in the culture medium as a 10× concentrated solution. 10 μ L suction costimulator to white 96 well plate, and immediately added to two duplicate 100 μ L of cell / compound mixture. The cells were stimulated at 37°C and 5% CO ₂ for 5.5 hours. After stimulation of the cells, each well was added 50 μ L BriteLifePlus reagent (Perkin Elmer), and to read the count Wallac EnVision (Perkin Elmer) measuring bioluminescence.

Any of the above assays can be scaled up for large-scale or high-throughput screening. Any of the above assay, IC ₅₀ values may be determined the subject compound. In a specific example, the IC ₅₀ value of the subject compound is determined using the method described in Example 1 and/or Example 2.

7. Pharmaceutical composition

The present invention provides a pharmaceutical composition, which comprises any one of the compounds described herein or a pharmaceutically acceptable salt thereof, and one or more pharmaceutically acceptable carriers or excipients.

"Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to the formulation and/or administration of the active agent and/or the absorption of the subject, and may be included in the composition of the present invention It is a substance that does not cause significant adverse toxicological effects on subjects. Non-limiting examples of pharmaceutically acceptable carriers and excipients include water, NaCl, normal saline, lactated Ringer's solution, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, Coatings, sweeteners, flavoring agents, salt solutions (e.g. Ringer's solution), alcohol, oil, gelatin, carbohydrates (e.g. lactose, amylose or starch), fatty acid esters, Hydroxymethyl cellulose, polyvinylpyrrolidone, and pigments, etc. Such preparations can be sterilized and, if necessary, can be combined with auxiliary agents that will not adversely react with or interfere with the activity of the compounds provided herein, such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, and effects. Osmotic pressure salts, buffers, colorants, and/or aromatic substances are mixed. Those with ordinary knowledge in the technical field of the invention will recognize that other medical carriers and excipients are suitable for use with the disclosed compounds.

These compositions may further contain one or more additional therapeutic agents as needed. Alternatively, the compounds of the present invention can be combined with one or more other treatment regimens [e.g. Gleevec or other kinase inhibitors, interferons, bone marrow transplantation, farnesyl transferase inhibitors, bisphosphonates, α- Phthaloimide, glutarimide, cancer vaccine, hormone therapy, antibodies, radiation, etc.] are administered to patients in need. For example, the therapeutic agent used for co-administration with the compound of the present invention or included in the pharmaceutical composition may be one or more other anti-cancer agents.

As described herein, the composition of the present invention comprises a compound of the present invention and a pharmaceutically acceptable carrier. As used herein, it includes any and all solvents, diluents or other vehicles, dispersion or suspension aids suitable for the specific dosage form required. Agents, surfactants, isotonic agents, thickeners or emulsifiers, preservatives, solid binders, lubricants, etc. Remington's Pharmaceutical Sciences, Fifteenth Edition, E.W. Martin (Mack Publishing Co., Easton, Pa., 1975) discloses various carriers for formulating pharmaceutical compositions and known techniques for their preparation. Unless any conventional carrier medium is incompatible with the compound of the present invention, for example, by causing any adverse biological effects or interacting with any other ingredients of the pharmaceutical composition in a harmful manner, its use is encompassed within the scope of the present invention. Some examples of materials that can be used as pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose , Ethyl cellulose and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppositories Wax; oils such as peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil; corn oil and soybean oil; propylene glycol such as ethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; Buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol and phosphate buffer, and other non-toxic compatible lubricants such as dodecyl Sodium sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweeteners, flavoring and aromatic agents, preservatives and antioxidants may also be present in the composition.

8. Formulations

The present invention also covers a type of composition comprising the active compound of the present invention and one or more pharmaceutically acceptable carriers and/or diluents and/or adjuvants (collectively referred to herein as "carrier" materials) and, Other active ingredients of time.

In specific embodiments, the present invention provides a pharmaceutical formulation for the treatment of cancer (especially the cancer described herein), which comprises a compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable carrier.

In a specific embodiment, the present invention provides a pharmaceutical formulation for the treatment of cancer selected from the group consisting of the treatable diseases described herein, which comprises the compound of the present invention or a pharmaceutically acceptable salt thereof and a pharmaceutically acceptable The carrier.

The active compound of the present invention can be administered via any appropriate route, preferably in the form of a pharmaceutical composition adapted to such route and in an effective dose for the intended treatment. The compounds and compositions of the present invention can be, for example, oral, mucosal, topical, rectal, for example, via inhalation spray via lung, or parenteral, including intravascular, intravenous, intraperitoneal, subcutaneous, intramuscular, and intrasternal With infusion technology, it is administered as a dosage unit formulation containing conventional pharmaceutically acceptable carriers, adjuvants, and vehicles.

The pharmaceutically active compound of the present invention can be processed in accordance with the conventional methods of pharmacy to produce It is used to administer medical preparations to patients (including humans and other mammals).

Regarding oral administration, the pharmaceutical composition may be in the form of, for example, tablets, capsules, suspensions or liquids. The pharmaceutical composition is preferably in the form of a dosage unit containing a specific amount of active ingredient.

Examples of such dosage form units are lozenges or capsules. For example, the appropriate daily dose for humans or other mammals can vary depending on the patient's symptoms and other factors, but can be determined again using routine methods.

The amount of compound administered and the dosage regimen of using the compound and/or composition of the present invention to treat disease symptoms depend on various factors, including the subject’s age, weight, sex and medical symptoms, disease type, disease severity, route of administration and Frequency and specific compound used. Therefore, the dosing regimen can vary widely, but can be determined routinely using standard methods. As previously mentioned, the daily dose can be administered once or divided into 2, 3, 4 or more administrations.

For therapeutic purposes, the active compound of the present invention is usually combined with one or more adjuvants, excipients, or carriers suitable for the designated route of administration. For oral administration, these compounds can be incorporated into lactose, sucrose, starch, cellulose alkanoate, cellulose alkyl ester, talc, stearic acid, magnesium stearate, magnesium oxide, sodium and sulfuric acid Calcium salt, gelatin, gum arabic, sodium alginate, polyvinylpyrrolidone, and/or polyvinyl alcohol are then tableted or encapsulated for convenient administration. Such capsules or lozenges may contain a controlled release formulation and may be provided as a dispersion of the active compound in hydroxypropyl methylcellulose.

In the case of skin symptoms, it is preferable to apply a topical preparation of the compound of the present invention to the affected area two to four times a day. Formulations suitable for topical administration include liquid or semi-liquid preparations suitable for penetrating the skin (such as liniments, lotions, ointments, emulsions, or pastes) and drops suitable for administration to the eyes, ears or nose. For topical administration, the active ingredient may account for 0.001% to 10% w/w of the formulation, for example, 1% to 2% by weight, although it may account for up to 10% w/w, but preferably no more than 5 % w/w, more for deployment 0.1% to 1% of the agent.

The compounds of the invention can also be administered via transdermal devices. It is better to use a kit and a patch with a variety of porous membrane types or solid substrates to achieve transdermal administration. In either case, the active agent is continuously passed through the membrane from the kit or microcapsule, and delivered to the adhesive permeable to the active agent in contact with the skin or mucous membrane of the recipient. If the active agent is absorbed through the skin, a controlled and predetermined flow of the active agent can be administered to the recipient. In the case of microcapsules, the encapsulating agent can also function as a membrane. The oil phase of the emulsion of the present invention can be composed of known ingredients by known methods.

Although this phase may contain only emulsifiers, it may contain a mixture of at least one emulsifier with fats or oils or with both fats and oils. Preferably, the hydrophilic emulsifier is used together with a lipophilic emulsifier as a stabilizer. More preferably, it contains both oil and fat. In short, emulsifiers with or without stabilizers constitute the so-called emulsifying wax, and the wax and oil together with the fat constitute the so-called emulsifying ointment base, which forms the oily dispersed phase of the emulsion formulation. Emulsifiers and emulsion stabilizers suitable for use in the formulations of the present invention include Tween 60, Span 80, cetearyl alcohol, myristyl alcohol, glyceryl monostearate, sodium lauryl sulfate, alone or together with wax The glycerol distearate, or other materials known in the technical field of the invention.

The selection of the appropriate oil or fat for the formulation is based on achieving the desired cosmetic properties. Since the solubility of the active compound in most oils that are likely to be used in pharmaceutical emulsion formulations is very low, the emulsion is preferably a non-greasy, non-staining and washable product that should have an appropriate consistency, so as to avoid pipes or other products. Leaking from the container. Can use linear or branched monobasic alkyl ester or dibasic alkyl ester, such as diisoadipate, isocetyl stearate, coconut oil acid propylene glycol diester, myristic acid Isopropyl ester, decyl oleate, isopropyl palmitate, butyl stearate, 2-ethylhexyl palmitate or a mixture of branched chain esters. They can be used alone or in combination depending on the required properties.

Alternatively, high melting point lipids can be used, such as white soft paraffin and/or liquid paraffin or its He mineral oil.

Formulations suitable for topical administration to the eye also include eye drops, in which the active ingredient is dissolved or suspended in a suitable carrier, especially an aqueous solvent for the active ingredient.

The active ingredient concentration present in such formulations is preferably 0.5 to 20%, advantageously 0.5 to 10%, especially about 1.5% w/w.

The formulations for injection administration may be in the form of aqueous or non-aqueous isotonic sterile injection solutions or suspensions. These solutions and suspensions can be prepared as sterile powders or granules using one or more of the carriers or diluents mentioned in formulations for oral administration, or using other appropriate dispersing or wetting agents and suspending agents. These compounds are soluble in water, polyethylene glycol, propylene glycol, ethanol, corn oil, cottonseed oil, peanut oil, sesame oil, benzyl alcohol, sodium chloride, tragacanth gum, and/or various buffers. Other adjuvants and administration modes are widely known in the technical field to which medical inventions belong. The active ingredient can also be dissolved with appropriate carriers (including saline, dextrose, or water), or with cyclodextrin (Captisol), co-solvent solubilization (i.e. propylene glycol) or micelle solubilization (i.e. Tween 80) The composition was administered by injection.

The sterile injectable preparation may also be a sterile injectable solution or suspension in a non-toxic injection-acceptable diluent or solvent, such as a solution in 1,3-butanediol. Among the acceptable vehicles and solvents, water, Ringer's solution, and isotonic sodium chloride solution can be used. In addition, sterile fixed oils are conventionally used as solvents or suspension media. For this purpose, any mild non-volatile oil can be used, including synthetic monoglycerides or diglycerides. In addition, the use of fatty acids (such as oleic acid) in the preparation of injections has been discovered.

For pulmonary administration, the pharmaceutical composition can be in the form of an aerosol or can be administered in an inhaler including dry powder aerosol.

Suppositories for rectal administration of drugs can be prepared by mixing the drug with appropriate non-irritating excipients such as cocoa butter and polyethylene glycol. These excipients are solid at room temperature, but are It is liquid at temperature, so it will melt in the rectum and release the medicine.

The pharmaceutical composition can be subjected to conventional medical operations such as sterilization and/or can contain conventional adjuvants, such as preservatives, stabilizers, wetting agents, emulsifiers, buffers, and the like. In addition, tablets and pills with enteric coating can be prepared. Such compositions may also contain adjuvants such as wetting agents, sweetening agents, flavoring agents, and fragrances. The pharmaceutical composition of the present invention comprises compounds of the formulae described herein or pharmaceutically acceptable salts thereof; selected from kinase inhibitors (small molecules, polypeptides, antibodies, etc.), immunosuppressants, anticancer agents, antiviral agents, Anti-inflammatory agents, antifungal agents, antibiotics, or additives for anti-vascular hyperplasia compounds; and any pharmaceutically acceptable carrier, adjuvant or vehicle.

The alternative composition of the present invention comprises a compound of the formulae described herein or a pharmaceutically acceptable salt thereof; and a pharmaceutically acceptable carrier, adjuvant or vehicle. Such compositions may optionally contain one or more additional therapeutic agents, including, for example, kinase inhibitors (small molecules, polypeptides, antibodies, etc.), immunosuppressants, anti-cancer agents, anti-viral agents, anti-inflammatory agents, and anti-fungal agents. Drugs, antibiotics, or anti-vascular hyperplasia compounds.

The term "pharmaceutically acceptable carrier or adjuvant" refers to a carrier or adjuvant that can be administered to a patient together with the compound of the present invention, which, when administered in a therapeutic amount of the compound in a sufficient dose, will not destroy the pharmacology Active and non-toxic. The pharmaceutically acceptable carriers, adjuvants and vehicles that can be used in the pharmaceutical composition of the present invention include, but are not limited to, ion exchangers, alumina, aluminum stearate, lecithin, and self-emulsifying drug delivery systems (SEDDS) (E.g. d- α -tocopherol polyethylene glycol 1000 succinate), surfactants used in pharmaceutical dosage forms (e.g. Tweens or other similar polymeric delivery matrix), serum proteins (e.g. human serum albumin), buffer substances (E.g. phosphate, glycine, sorbic acid, potassium sorbate), saturated vegetable fatty acid partial glyceride mixture, water, salt or electrolyte (e.g. protamine sulfate, disodium hydrogen phosphate, potassium hydrogen phosphate, sodium chloride , Zinc salt, silica gel, magnesium trisilicate), polyvinylpyrrolidone, cellulose substances, polyethylene glycol, sodium carboxymethyl cellulose, polyacrylate, wax, polyethylene polyoxypropylene blocking polymer , Polyethylene glycol and lanolin. Cyclodextrin (such as u-, P-, and y-cyclodextrin), or chemically modified derivatives (such as hydroxyalkyl cyclodextrin, including 2 and 3-hydroxypropyl cyclodextrin) or other soluble Derivatives can also be advantageously used to enhance the delivery of compounds of the formulae described herein.

The pharmaceutical composition can be administered orally in any orally acceptable dosage form, including, but not limited to, capsules, lozenges, emulsions and aqueous suspensions, dispersions and solutions. In the case of oral lozenges, carriers that are often used include lactose and corn starch. Lubricants such as magnesium stearate are usually also added. For oral administration in capsule form, useful diluents include lactose and dried corn starch. For oral administration of aqueous suspensions and/or emulsions, the active ingredients can be suspended or dissolved in the oil phase and combined with emulsifiers and/or suspending agents.

When necessary, specific sweeteners, flavoring agents and/or coloring agents can be added. The pharmaceutical composition may include a formulation using liposome or microencapsulation technology, various examples of which are known in the art to which the invention belongs.

The pharmaceutical composition can be administered via nasal aerosol or inhalation. Such compositions are prepared according to techniques known in the technical field to which the invention of pharmaceutical formulations belong, and can use benzyl alcohol or other appropriate preservatives, absorption promoters that enhance the availability of the body, fluorocarbons and/or other co-solvents Or the dispersant is prepared as a salt solution, examples of which are also known in the technical field of the invention.

9. Treatment kit

One aspect of the present invention relates to a kit for conveniently and effectively implementing the method or use according to the present invention. Generally, the pharmaceutical package or kit contains one or more containers filled with one or more of the ingredients of the pharmaceutical composition of the present invention. Such kits are particularly suitable for delivering solid oral forms such as lozenges or capsules. Such kits preferably contain a number of unit doses, and may also contain cards with dose orientations in the order of their intended use. need When necessary, memory aids can be provided, for example, in the form of numbers, letters, or other markings or using calendar inserts to mark the days on which the dose can be administered in the treatment plan. If necessary, related to such containers may be a notice in the form prescribed by a government agency that regulates the manufacture, use, or sale of pharmaceuticals, and the notice reflects the approval of the manufacturing, use, or sale organization of the human-administered drug.

The following representative embodiments contain more important information, examples and guidance, which can be applied to the implementation of the present invention in various specific examples and their equivalents. These embodiments are intended to help illustrate the present invention, but are not intended and should not be construed as limiting its scope. Indeed, after reviewing this document (including the subsequent examples and references to scientific and patent documents cited herein), in addition to those shown and described herein, various modifications of the present invention and many further specific examples, It will become obvious to those familiar with the technical field to which this invention belongs.

The contents of the cited references are incorporated herein for reference to help explain the state of the technical field of this invention.

In addition, for the purpose of the present invention, the chemical elements are identified in accordance with the Periodic Table of Elements (CAS version, Handbook of Chemistry and Physics, 75th Ed., on the cover). In addition, the general principles of organic chemistry and specific functional parts and reactivity are described in "Organic Chemistry," Thomas Sorrell, University Science Books, Sausalito: 1999 and "Organic Chemistry," Morrison & Boyd (3d Ed), two All the contents of the author are incorporated into this article for reference.

10. Synthesis process

The compound of formula I can be prepared by a person with ordinary knowledge in the technical field to which the invention belongs, following the techniques and procedures recognized in the technical field to which the invention belongs. More specifically, the compound of formula I can be prepared using the procedures, methods and examples set forth below. Those skilled in the technical field of the invention will recognize that the following flow can be changed Each step in the process to provide a compound of formula I. The reagents and starting materials are easily obtained by those with ordinary knowledge in the technical field to which the invention belongs. Unless otherwise specified, all substituents are as previously defined.

[Example]

The following are the abbreviations and their meanings used in this manual:

EtOAc/EA: ethyl acetate

DCM: Dichloromethane

ACN: Acetonitrile

THF: Tetrahydrofuran

DMSO: dimethyl subsulfate

MeOH: methanol

EtOH: ethanol

DMF: N,N-Dimethylformamide

DMA: N,N-Dimethylacetamide

DMF DMA: N,N-dimethylformamide dimethyl acetal

NCS: N-chlorosuccinimide

NBS: N-bromosuccinimide

NIS: N-iodosuccinimide

Pd-C: Palladium on carbon

LDA: lithium diisopropylamide

TFA: Trifluoroacetic acid

PTSA: p-toluenesulfonic acid

DIBAL-H: Diisobutyl aluminum hydride

LAH: Lithium Aluminum Hydride

PE: Petroleum ether

Py: pyridine

DPPA: Diphenylphosphoryl azide

CDI: 1,1-hydroxydiimidazole

TEA: Triethylamine

DIPEA: N,N-Diisopropylethylamine

DMAP: 4-(Dimethylamino)pyridine

EDCI: N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride

HOBT: 1-Hydroxybenzotriazole

TfOH: Trifluoromethanesulfonic acid

dppf: 1,1-ferrocene-diyl-bis(diphenylphosphine)

DAST: (Diethylamino) Sulfur Trifluoride

Pd ₂ (dba) ₃ : ginseng (dibenzylidene acetone) two palladium (0)

Boc: tertiary butoxycarbonyl

Ac: Acetyl

TMSI: Trimethylsilyl iodide

TBAI: Tetrabutylammonium iodide

PPh ₃ : Triphenylphosphine

dba: dibenzylidene acetone

BINAP: 2,2'-bis(diphenylphosphino)-1,1'-bis-naphthyl

MsCl: Methanesulfonyl chloride

TsCl: Tosyl chloride

DMAP: 4-Dimethylaminopyridine

LiHMDS: Lithium bis(trimethylsilyl)amide

NaHMDS: sodium bis(trimethylsilyl)amide

DMS: Dimethyl Sulfide

DAST: Diethylamine sulfur trifluoride

DME: Dimethoxyethane

DCE: Dichloroethane

NMR: Nuclear Magnetic Resonance

LC-MS: Liquid Chromatography Mass Spectrometry

ESI-MS: Electrospray Free Mass Spectrometry

GCMS: Gas Chromatography Mass Spectrometry

TLC: Thin Layer Chromatography

TCR: T cell receptor

BCR: B cell receptor

CARD: Apoptotic protease activation and recruitment domain

mM: millimolar

μM: micromolar

mL: microliter

ng: Naik

nM: Nemol

nm: Nano

IC ₅₀ : Half maximum inhibitory concentration

OD: Optical density

A. Biological Examples

Example 1. Raji/GLOSENSOR ^TM Compound screening of stable cell lines

This example provides an identification method capable of high-throughput screening/evaluation of MALT1 inhibitor activity in living cells. This identification method can be used, for example, to identify compounds that have effective in vivo activity on MALT1, identify MALT1 inhibitors that are more effective than other MALT1 inhibitors, and identify up to 70 to 80% (instead of 100%) MALT1 proteolysis MALT1 inhibitors that are part of the active (relative to complete) inhibitors, as well as the structure-function relationship between structurally related MALT1 inhibitors, etc.

The basic principle of the identification method described in this article is based on Fontan et al. [ J Clin Invest. 128(10): 4397-4412, 2018, incorporated herein for reference], which is used to detect MALT1 paratope in cells. It is a robust and sensitive method to prevent protease activity and the effect of any potential MALT1 inhibitor on MALT1 activity. This method is based on ^{the segmented luciferase system of GLOSENSOR TM} (Promega Corp., Madison, WI), which utilizes a genetically modified form of the firefly which is cut into two different domains by inserting a cAMP binding protein part Bioluminescent chimeric protein composed of luciferase. The binding of cAMP induces a configuration change, which rebuilds the functional luciferase protein to cause luminescence. Instead of inserting the cAMP-binding protein part, Fontan designed the reporter so that the configuration change is caused by the proteolytic cleavage induced by MALT1, so the luciferase activity will replace the endogenous MALT1 protease activity. More specifically, Fontan ^{uses the cleavage site sequence of Re1B (natural MALT1 substrate) in its GLOSENSOR TM} construct, so that the encoded luciferase is clearly activated by MALT1. ^{The GLOSENSOR TM} construct was tested in Raji lymphoma cells. These cells are easily transducible and suitable for this type of assay because they activate MALT1 immediately after stimulation with PMA and ionomycin (IO).

Therefore, in the assay described below, without antibiotics, grow the necessary number of Raji cells (for example, 5×10 ⁵ cells/mL) in a T75 culture flask overnight, so that the cells will reach 70 to 70 on the next day. 90% confluence. On the next day, cells of the optimal density were collected and counted for verification. After washing the collected cells in pre-warmed PBS (without Ca ²⁺ and Mg ²⁺ ), resuspend Raji cells in an appropriate resuspension buffer (including using Neon ^TM kit, Thermo fisher, # MPK1025) , The final density is about 1×10 ⁷ cells/mL.

Then, the addition of an appropriate capacity GLOSENSOR ^TM plasmid (for Raji cells, each well was 0.5 μ g) to the cell suspension. About EGFP control group, the 0.5 μ gEGFP plastids were added to the cell suspension.

By changing the pulse voltage, pulse width, and pulse number, test 24 different electroporation conditions, and select condition 16 in the following table (pulse voltage: 1400V; pulse width: 20 msec.; and pulse number: 2) for Electroporation. The same amount of EGFP plastid was also used to electroporate into Raji cells to evaluate transfection efficiency.

Two days after electroporation, Raji cells electroporated with EGFP constructs were collected, and the expression of EGFP was determined by FACS analysis. The efficiency of electroporation was determined by the gated FITC ^{+ ratio.}

Two days after electroporation, Raji cells electroporated with (or unused) GLOSENSOR ^TM constructs were collected, and then seeded into 96-well culture dishes at a density of about 10,000 cells per well. At 37°C, the cells were stimulated with PMA (200ng/mL)/ionomycin (1 μM ) for 2 hours to activate the MALT1 protease activity. Then, BRIGHT GLO ^TM (Promega Corp., Madison, WI) was added to each well to detect the luciferase signal. After PMA/IO stimulates the MALT1 activity, Raji cells expressing MALT1 were successfully electroporated ^{in the GLOSENSOR TM construct. After MALT1 cuts the GLOSENSOR TM} fusion protein to reconstitute the luciferase activity, it exhibits a strong luciferase signal.

In order to establish ^{Raji cells with the GLOSENSOR TM} construct firmly integrated into the genome, the electroporated Raji cells were collected two days after electroporation, and then plated in a 24-well culture dish with G418 medium. Select cells with a firmly integrated GLOSENSOR ^TM construct (which expresses the neomycin resistance gene). The cell culture medium was changed every three days, and the cells were centrifuged at 1,500 rpm for 5 minutes each time. After 13 to 14 days, pool all surviving cells (presumed to have a firmly integrated GLOSENSOR ^TM construct), freeze, and then use Bright-Glo matrix to evaluate luciferase activity.

By first diluting the pooled stable in Raji cells to a density of about 1 per 100 μ L cells, and then 100 μ L draw cell suspension (containing an average of one single cell) to each well of the 96 well plates, to obtain Single cell colonization of Raji cells with firmly integrated GLOSENSOR ^{TM constructs.} Let the cells grow for 3 to 4 weeks to grow into single cell clones.

After PMA/IO stimulation, the MALT1 activity of each single cell clone can be tested. Specifically, the cells can be plated in the wells of a 96-well culture plate at a density of about 20,000 cells/well, and then stimulated by adding 200ng/mL PMA plus 1 μM ionomycin. Add an equal volume of culture medium without PMA and IO to the control wells as a control group. Further, Raji cells without electroporation were used as a vehicle control, which produced negative signals under both stimulation and non-stimulation conditions. The cell clone with the measurement window greater than 7 is selected as the positive clone.

Further according to the present invention using the reference compound, test positive Raji such stable cell clones are measured IC ₅₀ via. Specifically, 40 μL of approximately 20,000 cell plates were cultured in each well of a 96-well culture plate. Meanwhile, the reference compounds (WO2015 / 181747 Example 10 disclosed in the embodiment RGT005-001) starting at a concentration of 200 μ M, 3-fold serial dilutions to a dose level of 10, then contacting the cell with the plate culture of each dose. After incubating at 37°C for 30 minutes, the cells were contacted with PMA/IO (or an equal volume of control medium) for 60 minutes to stimulate MALT1 activity, and then Bright Glo luciferase substrate was added to measure luciferase activity. The inhibition of MALT1 activity in the presence of the reference compound can determine the inhibition rate.

Any other test compound can be serially diluted in the same way, so that ^{the Raji stable cell line with integrated GLOSENSOR TM} construct can be contacted with different concentrations of potential MALT1 inhibitors to evaluate the IC ₅₀ value of each test compound, preferably high Flux form. MALT1 with high inhibitory activity can be identified using this assay.

The conditions and equipment used for representative analysis experiments are listed below.

Materials and media

1. Reagents and consumables

2. Equipment

3. Medium and solution

Raji's complete medium

Raji: RPM1640: 90%; FBS: 10%; Penicillin/Streptomycin: 1% Raji/GLOSENSOR ^TM complete medium

Raji: RPM1640: 90%; FBS : 10%; penicillin / streptomycin: 1%; G418: 1,500 μ g / mL

Freezing medium

FBS: 90%; DMSO: 10%

Stable cell line production procedure

1. Transfection/Electroporation

Day 1: Prepare the cells

Inoculate the required amount of cells (5×10 ⁵ cells/mL in a T75 culture flask) so that the cell confluence reaches 70 to 90% the next day. The medium does not contain antibiotics.

Day 2: Use Neon ^TM Transfection system (Thermo fisher#MPK5000) for electroporation

a. Collect the cell suspension. By mixing 20 μ L of the cell suspension and 20 μ L of trypan blue, cells were counted using Countstar machine. The cells were centrifuged at 1,000 rpm for 5 minutes.

b. Remove the supernatant and resuspend the cell pellet in pre-warmed PBS (without Ca ²⁺ and Mg ²⁺ ). The cells were centrifuged at 1,000 rpm for 5 minutes.

c. Resuspend the cell pellet in an appropriate resuspension buffer (included in the Neon ^TM kit, ThermoFisher, #MPK1025), Raji's final density is 1×10 ⁷ cells/mL.

d. Add the appropriate volume GLOSENSOR ^TM plasmid into the aforementioned cell suspension (for Raji cells, each well was 0.5 μ g). For EGFP group, respectively, added 0.5 μ g of plasmid EGFP to cell suspension.

e. In the mass before transfection, the culture containing no antibiotic was drawn into 24-well culture plates, each well of 500 μ L. Preheat the culture plate at 37°C for a while.

f. Use condition 16 (pulse voltage: 1400V; pulse width: 20 msec; pulse number: 2) Raji was electroporated ^{with GLOSENSOR TM mass.} The same amount of EGFP was also electroporated into Raji cells to evaluate the transfection efficiency. Note: A total of 24 transfection conditions were tested (see below), and condition 16 was selected as the optimal condition.

Day 4: Evaluation of transfection/electroporation efficiency

Collect cells in EGFP electroporated wells and perform FACS. The transfection efficiency was evaluated by the gated FITC ^{+ ratio.}

Day 4: Assessment of transient transfection signal

Collect cells transfected with or without GLOSENSOR ^TM. The cells were seeded into 96-well culture plates with 10,000 cells per well. The cells were stimulated with PMA (200ng/mL)/ionomycin (1 μM ) at 37°C for 2 hours, and then Bright Glo was added to detect the luciferase signal.

Day 4 to Day 17: Antibiotic treatment

a. On the 4th day, transfer the cells in the 24-well culture dish to the T25 culture flask. Add 1,500 μ g / mL G418, Bishi give Raji / GLOSENSOR ^TM clones are stabilized.

b. Replace the cell culture medium by centrifugation at 1,500 rpm for 5 minutes every three days.

c. On the 17th day, collect the pooled stable cells. It was frozen and Bright-Glo was used to evaluate the signal of pooled cells.

2. Preparation of single cell clones

a. Dilute the pooled stably transfected cells to 1 cell/100 μL .

b. Pipette 100 μ L of the cell suspension into 10 96-well plates.

c. Let the cells grow for 3 to 4 weeks. Mark the colony gradually formed from a single cell.

3. Use Bright-Glo to initially identify Raji/GLOSENSOR ^TM Positive clones

a. Observe the cell status of each stable clone under a microscope. Roughly evaluate the cell density of each clone.

b. Place the cells were plated in 96-well culture plate (Corning # 3610) in, 90 μ L / hole of about 2 × 10 ⁴ cells.

C. stimulus'argument group, 10 μ L of 200ng / mL PMA plus 1 μ M ionomycin was transferred to 96-well culture plate.

About unstimulated group, the system 10 μ L culture was transferred to each well of 96-well culture plate. Un-electroporated Raji cells were used as the vehicle control group, which should produce negative signals under both stimulation and unstimulation.

d. Incubate the culture plate for 2 hours in a 37°C, 5% CO _{2 incubator.}

E. Add Bright-Glo 100 μ L / hole of the culture dish.

f. At room temperature, shake the culture plate (~300rpm) for 5 minutes on the culture plate shaker.

g. Read the luminescence signal in the Envision Micro Disk Analyzer (96-well USL).

h. Processing data. Check the test window of each selected plant between the stimulated and non-stimulated groups. Select clones with a test window greater than 7 as positive clones.

4. Use the IC of the reference compound RGT005-001 ₅₀ Confirmation of positive clones

a. Cells were seeded into 96-well culture plate (Corning # 3610) per well in 40 μ L, 20,000 cells.

b. Compound preparation:

i) The RGT005-001 (dissolved in DMSO stock solution of 10mM) diluted to 200 μ M (1 μ L stock solution was added to 49 μ L DMSO to obtain 200 μ M) using DMSO.

ii) at 200 μ M RGT005-001 as the highest dose. Then 3-fold dilutions (the highest dose of 10 μ L of solution with mixing 20μL DMSO) into 10 dose levels.

iii) preheating compounds were serially diluted (2 μ L of the compound solution with mixing 198 μ L broth) to give compound 2 × broth.

iv) in 96 well plates in 40 μ L of compound solution treated cells.

v) Shake at 300 rpm for 2 minutes, and incubate the assay plate for 30 minutes _{at 37°C and 5% CO 2.}

vi) respect to volume / low for the control group, based adding 40 μ L broth (containing 1% DMSO) to the culture dish.

c.30 minutes, 20 μ L 5 × PMA / ionomycin were transferred to 96-well culture plate.

D. volume control system is added 20 μ L PMA / ionomycin to the culture dish. Low volume-based control group was added 20 μ L broth was added to the culture dish.

e. Incubate the culture plate in an incubator at 37°C and 5% CO _{2 for 1 hour.}

F. Adding Bright-Glo 100 μ L / hole of the plate to the assay.

g. At room temperature, shake the plates (~300rpm) for 5 minutes on a culture plate shaker.

h. Read the luminescence signal in the Envision Micro Disk Analyzer (96-well USL).

i. Use the equation to process the data to obtain the inhibition rate: (reading of high-volume control wells-reading of compound-treated wells)×100%/(reading of high-volume control wells-reading of low-volume control wells). For different concentrations of compounds, fitting inhibition rates. Statistical analysis using graphics software (GraphPad prism) in the nonlinear analysis yielded IC _50.

5. Mycoplasma detection

Use MYCOALERT ^TM PLUS mycoplasma detection kit (order information: Lonza, LT07-710) for mycoplasma detection, and strictly follow the instructions provided by the supplier for the determination.

At Raji/GLOSENSOR ^TM Procedures for screening compounds on cell lines

Inoculate cells in 96-well culture plates:

Cells were seeded into 96-well culture plate (Corning # 3610) per well in a total of 2 × 10 ⁴ cells (40 μ L). Note: Record the number of cells and cell viability each time.

Compound serial dilution

Regarding the reference compound RGT005-001:

a. with DMSO RGT005-001 (dissolved in DMSO stock solution of 10mM) diluted to 200 μ M (1 μ L stock solution was added to 49 μ L DMSO to give a 200 μ M).

B. then serially diluted with DMSO to a dose level of 10 (10 μ L with the highest dose of 20 L DMSO mixed solution) 3 times.

c. Make the solution of the compound (100 ×) was mixed with the preheated liquid culture, to give a solution of compound 2 × (so that the 2 μ L of compound solution was mixed with 198 μ L culture). The final concentration of DMSO is 1% (2×).

About the test compound:

a. with DMSO of the test compound (dissolved in DMSO stock solution of 10mM) diluted to 2mM (3 μ L stock solution to 12 μ L DMSO was added to give a 2mM).

B. Then 2mM with DMSO of the test compounds, diluted into 10 continuously repeated dose levels (5 μ L was the highest dose mixed with 10 μ L DMSO) 3 times.

C. The culture was mixed using preheated solution of compound (100 ×), to give a solution of compound 2 × (so that the 2 μ L of compound solution was mixed with 198 μ L culture). The final concentration of DMSO is 1% (2×).

Use compound to Raji/GLOSENSOR ^TM Cell pretreatment

a. at varying concentrations below (Platemap) the compound treated Raji / GLOSENSOR ^TM, each well was 40 μ L.

b. Shake the culture plate at 300 rpm for 2 minutes, and incubate the culture plate for measurement _{at 37°C and 5% CO 2 for 30 minutes.}

C. About volume / low volume control, the system 40 μ L broth (containing 1% DMSO) was added to the culture dishes.

PMA/ionomycin stimulation for 1 hour

a. Preparation of a mixture of pre-culture was 1 μ g / mL PMA plus ionomycin of 5 μ M (to 7984 μ L broth was added 8 μ L PMA and ionomycin 8 μ L).

b. After treating the compound for 30 minutes, transfer 20 μ of the LPMA/ionomycin mixture to a 96-well culture dish (anticipated positive control).

C. About low for the control group, the culture system was added 20 μ L was added to all wells.

d. Incubate the culture plate in an incubator at 37°C and 5% CO _{2 for 1 hour.}

Luciferase signal detection and data analysis

a. Add 100 μ L / hole using Bright-Glo assay to culture dishes.

b. At room temperature, shake (~300rpm) on the culture dish on the culture dish shaker (~300rpm) for 5 minutes.

c. Read the luminescence signal in the Envision Micro Disk Analyzer (96-well USL).

d. Use equation to process data into inhibition rate: (reading of high volume control well-reading of compound treatment well) × 100%/(reading of high volume control well-reading of low volume control well). For different concentrations of compounds, fitting inhibition rates. Statistical analysis using graphics software (GraphPad prism) in the nonlinear analysis yielded IC _50.

The Glosensor dose-response diagram of the exemplary compound of the present invention is presented in FIG. 1.

Example 2. Study on the efficacy of compounds produced by IL-2 in Jurkat cells

This example demonstrates that the subject compound affects IL-2 production in Jurkat cells. At the same time, ELISA can be used to test the ability of the test compound to affect IL-2 production in the same system.

Jurkat cells were selected for this analysis because they are immortal human T lymphocytes capable of producing interleukin 2 (IL-2). ^{T Cell TRANSACT™} reagent (Miltenyi Biotec) can be used to stimulate Jurkat cells to produce IL-2. This reagent has been developed as a ready-to-use reagent that can activate and expand human T cells via CD3 and CD28. Its polymeric nano-matrix structure composed of colloidal polymeric nano-matrix combined with anthropomorphic CD3 and CD28 agonists can ensure gentle and effective activation of T cells while maintaining high viability.

The Jurkat cells stimulated to produce IL-2 can be centrifuged, and the IL-2 containing supernatant can be collected, and then the amount of IL-2 produced can be measured using an anti-IL-2 antibody using a standard ELISA assay. The presence of MALT1 inhibitors reduces the amount of IL-2 produced by Jurkat cells, and this analysis method can be used to quantitatively measure the IC ₅₀ value of each MALT1 inhibitor.

The following is an exemplary analysis experiment procedure provided in this article.

Materials and equipment:

1. Reagents:

2. Equipment and supplies:

3. Procedure

Jurkat cell thawing and inoculation procedures:

Cell growth medium: RPMI1640 (ATCC, Cat #30-2001); 10% FBS (Gibco, catalog number 10091148); 1% PenStrep (Gibco, catalog number 15140); and Beta-ME (Gibco, catalog number 21985023) ).

1. Subculture

This cell line is usually diluted at a ratio of 1:3 twice a week.

1) Maintain cell culture by adding fresh medium/changing medium or centrifugation, and then ^{resuspend at 3 to 4×10 5} cells/mL.

2) When the cell concentration/density reaches 1×10 ⁶ cells/mL, the cells are subcultured. The cell concentration is controlled not to exceed 3×10 ⁶ cells/mL.

3) Renew the culture medium every 2 to 3 days.

2. Replace the fresh culture medium before the test day

3. Inoculate cells in a 96-well culture dish (day 1)

1) Collect the cells by centrifugation and discard the cell growth medium. Calculate the number of cells, and calculate the total number of cells required for the determination.

2) Dilute the cell suspension with an appropriate volume of cell culture medium.

3) The cell suspension is then dispensed into a sterile disposable kit.

4) and then 1 × 10 ⁵ cells / well (50 μ L / well) of cell suspension was transferred to a 96 well plate (cell culture plate: corning-3788) in.

5) Incubate the culture plate for 1 hour in a humid air, 37°C, 5% CO _{2 incubator.}

Compound and Transact preparation procedure (Day 1):

1. Preparation and addition of compound and DMSO:

Compound serial dilution (source plate)

1) reacting a compound dissolved in 100% DMSO to a concentration of 10mM, then diluted with DMSO to 1mM: 5 μ L 10mM compound +45 μ L DMSO = 50 μ L 1mM Compound A was added to the column.

2) from about 40 μ L DMSO was added to all wells B1-H10. Then transferred via a 20 μ L to 40 μ L DMSO compound, a pipetteman serially diluted compound (8 grade, diluted 3-fold, B2 to H9). Through this method, the concentration of a series of compounds diluted in DMSO is obtained.

2×Compound dosage preparation [Middle plate: Corning-3599]:

1) to pipetteman added 95 μ LRPMI 1640 medium (RPMI1640 + 10% FBS + 1 % PS + 0.055mM 2-ME) to the hole 96 in the intermediate plate 1, then 5 μ L of various compounds from the source culture dishes Transfer to this intermediate plate 1 (A2 to H9) and mix;

2) In the pipetteman 5 μ L DMSO is transferred from the source to this intermediate plate culture tray 1 (A1 to H1, A10 to H10) and mixed thoroughly.

3) add to pipetteman 240 μ L RPMI 1640 medium (RPMI1640 + 10% FBS + 1 % PS + 0.055mM 2-ME) to the intermediate tray 96 2 (A1 to H10), and then to pipetteman 10 μ L will such a compound or DMSO are transferred into an intermediate tray from the intermediate tray 2 (A1 to H10) and thoroughly mixed to obtain a final compound concentration of 2 ×.

4) Add 50 μ L / hole working concentration (working concent solution to the cell culture plate (Corning-3788) than n) DMSO compound. The final concentration of compound 1 μ M, cell culture and final DMSO concentration of 0.1%. Then incubate the culture plate in a _{5% CO 2} incubator at 37°C for 0.5 hours.

2. Transact ^TM reagent: store at 4 ℃

1) The ^{working solution of the Transact TM} reagent should be freshly diluted from the stock solution with RPMI 1640 culture medium (RPMI1640+10% FBS+1% PS+0.055mM Beta-ME) before use. The sample is not diluted.

2) Add 100 μ L / 2 × Transact hole in each well.

3) Incubate for 24 hours in a _{5% CO 2 incubator at 37°C.}

Supernatant collection procedure

Sample collection (day 2)

1) After incubating in a 5% CO ₂ incubator at 37°C for 24 hours, centrifuge the cell culture plate at 1,000 rpm for 10 minutes. Collecting 100 μ L / well supernatants, followed by IL-2 ELISA assay. The supernatant can be injected at -80°C for IL-2 ELISA assay on the next day. If necessary, the supernatant can be diluted 3 times to ensure that the measurement does not exceed the linear range of the IL-2 standard curve.

IL-2ELISA program (day 2):

Coat the culture plate on day 1:

1) diluted capture antibody coated microtiter wells (each well 100 μ L) in coating buffer. Use the lot specific instructions/test certificate as a guide for recommending antibody coating dilutions. The culture plate was sealed and incubated at 4°C overnight.

IL-2 ELISA on day 2:

300μL/孔之洗滌緩衝液洗滌三次。最後一次洗滌後，將培養盤倒置並於吸水紙上吸乾以任何殘留之緩衝液。 1) Suction the holes to

Washed with 300 μ L / hole of the solution washed three times with buffer. After the last wash, turn the culture plate upside down and blot dry on absorbent paper to remove any remaining buffer.

200μL/孔之測定稀釋劑封阻培養盤，然後於室溫培育1小時。 2) to

Determination 200 μ L / hole of diluent blocker culture plate, and incubated at room temperature for 1 hour.

3) Repeat the suction/washing as in step 2.

4) Prepare standard and sample diluents in the test diluent.

Preparation of IL-2 standard stock solution: Add 1 mL of deionized water to a vial (235ng/vial) to achieve a stock solution concentration of 235ng/mL. The standard stock solution is equally divided into 50 μ L / vial, and then frozen at -80 ℃.

The STD curve is diluted with stock solution on the day of measurement, with the highest 500pg/ml, 2-fold dilution, and 8 doses (including 0pg/ml). After dilution, the STD 100 μ L / hole ELISA plates were transferred to the (line 12).

5) suction 100 μ L of each standard, sample and control to the appropriate wells. Seal the culture plate and incubate at room temperature for 2 hours.

6) Repeat the suction/washing as in step 2, but wash it five times in total.

7) Add 100 μ L working object detection (detection antibody reagent + SAv-HRP) to each well. The culture plate was sealed and incubated at room temperature for 1 hour.

8) Repeat the aspiration/washing as in step 2, but wash a total of seven times.

Note: In this final washing step, the wells are immersed in washing buffer for 30 seconds to 1 minute for each wash.

9) 100 μ L substrate solution was added to each well. Incubate the culture plate (unsealed) for 30 minutes at room temperature in the dark.

10) Add 50 μ L of stop solution to each well.

11) Within 30 minutes of stopping the reaction, read the absorbance at 450nm. If wavelength correction is possible, subtract the absorbance at 570nm from the absorbance at 450nm.

data analysis:

Calculate the inhibition percentage (%) of the concentration of each compound based on and relative to the signals in the wells of HPE and ZPE contained in each assay culture plate. Use HPE wells as 0% inhibition, and except for DMSO (final concentration = 0.1%), use ZPE wells without any compound as 100% inhibition. Plot the concentration of the test compound and the% inhibition value, and use the three-parameter logarithmic dose-response equation to determine the compound concentration (IC ₅₀ ) required for 50% inhibition. In each experiment, the result variable value (IC ₅₀ ) of the reference peptide/compound was evaluated as a benchmark for quality control. If the result variable value is within 3 times of the expected value, the experiment is considered acceptable.

In Table 1 below, cell data obtained from exemplified compounds are listed. For values less than or equal to 100nM, the IC ₅₀ value is expressed as "++++"; for values less than or equal to 500nM, "+++"; for values less than or equal to 1 μM , "++"; greater than The value of 1 μ M, "+".

Figure 2 presents the IL-2 Jurkat cell Transact dose-response graph of the exemplified compounds of the present invention.

Example 3. Inhibitory activity against Treg and Th17

CD4 ⁺ T cells play a key role in regulating the immune system to resist infection by pathogens including bacteria or viruses and cancer. The unique function of the CD4 ⁺ T cell compartment depends on the adjustable balance between various T cell subsets, among which Th17 cells and regulatory T cells (Tregs) play an important role in regulating autoimmunity and cancer. The delicate balance between Th17 cells and Treg cells is not only essential for maintaining a healthy and functioning immune environment, but also for the treatment of diseases. In other words, the purpose of offsetting the Th17/Treg balance may be effective in treating diseases related to the Th17/Treg balance and the disintegration of the steady state in the body.

Th17 cells are necessary for the host to defend against pathogens to maintain life, and are involved in initiating autoimmune diseases and cancer, while Tregs are necessary for self-tolerance and defense against autoimmunity, and are often correlated with cancer progression. This example demonstrates that, based on the measurement and analysis of the inhibitory function of the subject compound on Th17 or Treg function (including the expansion of differentiated Th17 or Treg cells), the compound of the present invention has a distinctive inhibitory effect on Th17 and Treg T cells.

A. Th17 assay-7 days culture to produce polarized human Th17-like cells

By ^{resuspending CD4+} naive T cells in a mixture containing anti-CD28 costimulatory antibodies (to provide a "second" activation signal), a mixture of cytokines (such as IL-6 (necessary for Th17 differentiation), IL-1 β (to promote Th17 Differentiation), TGF- β 1 (necessary for Th17 differentiation), IL-23 (necessary for supporting Th17 phenotype)), and anti-IL-4 (inhibition of initial T cell polarization into Th2 cells) and anti-IFN- γ (inhibition of initial T cells are polarized into Th1 cells) neutralizing antibody culture medium, and then the cells are plated in tissue culture wells pre-coated with anti-CD3 antibody (providing the "first" activation signal), and then cultured in vivo for 7 days Differentiate into human Th17-like cells. The differentiated Th17-like cells were collected and plated in a mixture of cytokines co-stimulated with anti-CD3 and anti-CD28 to measure the IL-17A production in the presence and absence of compounds in decreasing concentrations, so as to evaluate the test compound's inhibition of IL- The ability to produce 17A and/or Th17 function.

LANCE ^® (Lanthanide Chelate Exeite) TR-FRET technology was used to evaluate the production of IL-17A with a standard curve. In short, the LANCE ^® TR-FRET measurement is a combination of Time Difference Fluorescence (TRF) and Fluorescence Resonance Energy Transfer (FRET) for detection and quantification in a high-throughput format. It uses a long-lived fluorescent lanthanide chelate (Ulight), which allows the delay between excitation and emission to be measured. The Ulight fluorophore was conjugated with the first anti-IL-17A antibody (ie, ULight-labeled anti-hIL17A antibody), and the second anti-IL-17A antibody labeled ^{with LANCE ® Europium W-1024-ITC chelate (} That is, Eu-labeled anti-hIL17A antibody) is used together. When the two antibodies bind to IL-17A at the same time, the excitation of the Eu moiety results in the emission of detectable light from the Ulight fluorophore via TRF-FRET. The amount of IL-17A in this reaction is proportional to the detectable signal. Use a standard curve generated from a series of IL-17A samples with known concentrations to measure the IL-17A concentration in the test sample.

For FACS assays, PMA/ionomycin is used to stimulate Th17-like cells to promote the production of cytokines, and brefeldin-A is used to interrupt intracellular transport and retain the synthesized cells in the cells Interleukin, for FACS using fluorescently labeled anti-IL-17A Ab.

Reagents:

Equipment and supplies:

Differentiation assay experimental procedure

1. sterile PBS, anti-CD3 antibody was diluted to 10 μ g / mL.

2. L was diluted to 300 μ anti-CD3 antibody-coated 24-well flat bottom tissue culture plate. The culture plate was sealed and incubated at 4°C overnight for antibody cross-linking.

3. The next day, washed twice with culture plate coated to 1,000 μ L of sterile PBS.

4. The vial 1 initial (CD4 ^+, CD45RA ⁺⁾ T cells (AllCells) re-suspended in anti-CD28 antibody (1 μ g / mL), IL-6 (10ng / mL), IL-1 β (10ng / mL ), TGF- β 1 (5ng / mL), IL-23 (10ng / mL), and anti-IL-4 (10 μ g / mL) and anti-IFN- γ (10 μ g / mL ) antibodies and the like In X-Vivo 15 culture medium, the density is 1.0×10 ⁵ cells/mL.

QC criteria for donor selection:

PBMC QC: Fresh; CD3 ⁺ , 40-60%; CD4 ⁺ /CD8 ⁺ , 1.5-2.

CD4 ⁺ Initial T cell QC: fresh;

Sorting method: double negative selection

Living cells: > 95%

CD3 ⁺ CD4 ⁺ CD45RA ⁺ : > 95%

5. Add 1 mL of T cell suspension to a 24-well culture plate pre-coated with anti-CD3 antibody. Culture the cells in an incubator at 37°C and 5% CO _{2 for 7 days.}

6. On the 7th day:

Cells were collected for IL-17A FACS assay, and the cells were seeded in 96-well plates, such as for test compound (total test system 150 μ L) of the following:

1) of the resuspended cells are seeded in culture dishes precoated with anti-CD3 antibodies of the 96 X-vivo culture medium (2 × 10 ⁴ cells / well, 100 μ L / hole, 2 × 10 ⁵ cells / mL ).

2) Incubate in an incubator for 1 hour.

3) In the experiments the differentiation process used to maintain all of Th17 phenotype and cytokine antibodies (96-well culture plate in the presence of 50 μ L per well) using successively different concentrations to the culture broth prepared in X-vivo The diluted compound (3-fold serial dilution, from 10,000 nM to 4.57 nM) treated cells for 72 hours (37°C, 5% CO ₂ ).

4) on volume control wells, and antibody-based preparation of a cytokine in the X-vivo culture medium of 0.2% DMSO (96 well plates each well 50 μ L).

5) For the low volume control wells containing no antibody prepared based cytokine in the culture medium of 0.2% DMSO (96 well plates each well 50 μ L) in X-vivo.

6) After 72 hours, supernatant was collected from 50 μ L for each well was measured by HTRF, and test cells in FACS assay.

FACS measurement procedure

1) Preparation of PMA (final concentrations in the X-vivo culture medium without antibody to the cytokine in: 50ng / mL) and ionomycin (final concentration: 1 μ g / mL) and eBioscience ^TM Brefeldin A , Used for all holes.

2) adding PMA 50 μ L / hole of / ionomycin (P / I) and eBioscience ^TM Brefeldin A solution (1,000 ×) in all wells. Stimulate the cells for another 6 hours.

3) Collect cells for FACS determination.

4) Wash the cells once with 1 mL of PBS, and centrifuge at 300 g for 5 minutes.

5) Resuspend the cells in 1 mL PBS. The reaction was added fluorescent dye (prepared previously) -APC Cy7 1mL cell suspension to 1 μ L of reconstituted, and mixed thoroughly. Incubate for 20 minutes at room temperature in the dark, and centrifuge at 300g for 5 minutes.

6) Wash the cells once with 1 mL of PBS, and centrifuge at 300 g for 5 minutes.

7) The cells were resuspended in 100 μ L staining buffer. At room temperature, in the dark, the ^{cells were stained with CD4 +} -PE antibody for 30 minutes, and centrifuged at 300 g for 5 minutes.

8) The cells were washed twice in 500 μ L staining buffer, centrifuged at 300g for 5 minutes.

9) 4 ℃, the cells were resuspended in 100 μ L fixative buffer for 20 min.

10) Wash the cells twice in 1*Perm/Wash buffer.

11) Anti-IL-17A prepared in 1*Perm/Wash buffer, add 100 μL of anti-IL-17A in the dark at 4°C for 40 minutes.

12) Wash the cells twice with 1*Perm/Wash buffer and resuspend them in the staining buffer, and then perform flow cytometry analysis.

13) Once staining is complete, obtain cells for data analysis.

14) Describe the percentage of IL-17A positive cells.

IL-17A LANCE ^® Measurement procedure

1. The buffer was prepared 1 × Ultra HiBlock: Add 400 μ L 5 × HiBlock buffer to a 1,600 μ L of water.

2. Preparation of hIL-17A standard dilutions of the analyte: at H ₂ O and then configured to generate a 10 μ g / mL of solution. Preparation of 3-fold serial dilutions of the standard, the maximum concentration of 1.00E-06g 15 μ L of / hIL-17A, the minimum concentration mL of 3.00E-12g 15 μ L of / mL of hIL-17A (between each standard dilution Replace the straw head).

3. Preparation of anti hIL17A 4 antibody (1.2nM) × MIX Eu + ULight marker of the labeled anti-antibody hIL17A (12nM): Add 500nM Eu 1.32 μ L of labeled antibody to the anti hIL17A 13.2 μ L of the marker 500nM Ulight Anti-hIL-17A antibody.

4. In the white 384 culture dish:

A: Add 15 μ L of each standard dilution or analyte 15 μ L sample was centrifuged for 1 min at 1,000rpm.

B: adding 5 μ L 4 × MIX Eu labeled anti-hIL-17A of antibody (final 0.3nM) + Ulight of labeled anti-hIL-17A antibody (final 3nM), centrifuged for 1 min at 1,000rpm.

5. Incubate for 120 minutes at RT (room temperature).

6. Use LANCE TRF Laser to read (in TR-FRET mode).

B. Treg differentiation-6 days culture to produce polarized human Treg cells

Using the CellXVivo Human Treg Cell Differentiation Kit from R&D Systems, Inc. (Minneapolis, MN) (catalog number CDK006), according to the manufacturer’s recommendations, after 6 days of in vivo cultivation, the CD4 ⁺ initial T cells were differentiated into human Tregs cell. The kit contains optimized proteins and reagents to drive the initial CD4 ⁺ T cells to efficiently differentiate into FoxP3 ⁺ CD25 ⁺ Treg cells. After 6 days of culture, the CD4 ⁺ CD25 ⁺ and FOXP3 ⁺ expressions of differentiated Tregs were confirmed by FACS assay.

Specifically, for FACS assays, in order to express CD4 and CD25, Tregs are conjugated with phycoerythrin (PE). The fluorophore of anti-CD4 antibody (CD4 ⁺ -PE) is conjugated with isophycocyanin (APC) The anti-CD25 antibody (CD25 ⁺ -APC) fluorophore stains, then fixes and permeabilizes the cells for the use of anti-FOXP3-FITC antibody to detect the performance of FOXP3. The stained cells were analyzed by flow cytometry to determine the percentage of ^{FOXP3 + positive cells.}

At the same time, the differentiated Tregs were collected, and then plated in a mixture of cytokines co-stimulated with anti-CD3 and anti-CD28, and the IL-10 production of Tregs was measured in the presence and absence of compounds at decreasing concentrations, so as to evaluate the inhibition of IL by the test compounds -10 ability to produce. The IL-10 ELISA kit (catalog number 430604) from Biolegend (San Diego, CA) was used to measure the production of IL-10 according to the manufacturer's experimental procedure.

Materials and equipment:

Reagents:

Equipment and supplies:

Differentiation analysis program:

1. Reconstitute buffer 1 with 125 μL, and reconstitute human Treg reagent 1 and human Treg reagent 2 to produce a 400× stock solution.

2. Reconstitute buffer 2 with 125 μL and reconstitute human Treg reagent 4 to produce a 400× stock solution.

3. Add 125 μ L each of human Treg reagent 1,2, and 4 (400 × stock solution) and 50 μ L of human Treg reagent 3 (1,000 ×) to 49.6mL of X-VIVO 15 culture medium without serum.

4. L was diluted to 300 μ anti-CD3 antibody-coated 24-well flat bottom tissue culture plate. The culture plate was sealed and incubated at 4°C overnight for antibody cross-linking.

The first two days, washed twice with buffer tray 1,000 μ L of culture solution was washed with a coating.

6. Resuspend 1 vial of initial T cells (CD4 ⁺ , CD45RA ⁺ ; AllCells) in the X-Vivo 15 culture medium containing the human Treg reagent prepared in step 3 at a density of 1×10 ⁵ cells/mL.

QC criteria for donor selection:

PBMC QC: Fresh; CD3 ⁺ , 40 to 60%; CD4 ⁺ /CD8 ⁺ , 1.5-2.

CD4 ⁺ Initial T cell QC: fresh;

Sorting method: double negative selection

Living cells: > 95%

CD3 ⁺ CD4 ⁺ CD45RA ⁺ : > 95%

7. Add 1 mL of cells to a 24-well culture plate pre-coated with anti-CD3 antibody.

8. Incubate the culture plate in an incubator at 37°C and 5% CO ₂ for 6 days.

9. On day 6, cells were harvested for FACS assay measuring Foxp3 ⁺ CD25 ⁺ CD4 ⁺ T cells, and the cells were seeded in 96-well culture plate for test compound (total of 150 μ L of test systems):

1) and the resuspended cells are seeded in culture dishes precoated with anti-CD3 antibodies of the 96 X-vivo culture medium (1 × 10 ⁴ cells / well, 100 μ L / hole, 1 × 10 ⁵ cells / mL ).

2) Incubate in an incubator (37°C, 5% CO ₂ ) for 1 hour.

3) in the presence of reagent preparation of human Treg differentiation Protocol Step 3 (50 μ L per well to 96 well plate), was used for the preparation of X-vivo culture of different concentrations of compounds were serially diluted (3 fold serial Dilute, treat cells from 10,000 nM to 4.57 nM for 72 hours (37°C, 5% CO ₂ ).

4) on volume control wells, containing at step differentiation Protocol 3 of the prepared 0.2% DMSO (50 μ L / prepared hole to a 96-well plate) X-vivo culture medium of human Treg the reagent.

5) Regarding the amount of the low control wells, 0.2% DMSO (50 μ L / hole to 96 well plates) Preparation of X-vivo culture medium of human Treg agent 3 prepared in the step of containing no differentiation of Protocol.

6) After 72 hours, the supernatant was collected 120 μ L, IL-10 production using ELISA assay tests of Treg cells, and the cells were collected for assay by FACS to confirm that in the CD4 ⁺ CD25 ⁺ FoxP3 ⁺ on the expression of Tregs ( Only high-volume control group and low-volume control group) and CTG measurement (compound wells plus high-volume control/low-volume control).

FACS assay procedure:

1. Wash the cells once with 1 mL of PBS and centrifuge at 300 g for 5 minutes.

2. Resuspend the cells in 1 mL PBS. Fluorescent dye The reaction was added 1 μ L of the reconstituted (previously prepared) solution and mixed well to -APC Cy7 1mL cell suspension. Incubate for 20 minutes at room temperature in the dark, and centrifuge at 300g for 5 minutes.

3. Wash the cells once with 1 mL of PBS and centrifuge at 300 g for 5 minutes.

4. The cells were resuspended in 100 μ L staining buffer. In the dark at room temperature, and phycoerythrin (PE) anti-CD4 antibody bonded (CD4 ⁺ -PE) of the fluorophore and iso phycocyanin (APC) anti-CD25 antibody bonded (CD25 ⁺ -APC) of fluorescence And stain the cells for 30 minutes.

The cells were washed twice in staining buffer in the 500 μ L, centrifuged at 300g for 5 minutes.

6.4 ℃, the cells were resuspended in 500 μ L fixation / permeabilization buffer (4 ×, diluted with fixation / permeabilization of dilution) for 30 minutes. Centrifuge at 300g for 5 minutes at 4°C.

7. Add 500 μ L 1 × permeabilized wash buffer (diluted in ddH ₂ O from 10 ×) in resuspension. Centrifuge at 300g for 5 minutes at 4°C.

8. Add 100 μ L of anti-FOXP3-FITC antibody premix (prepared in permeabilization 1 × wash buffer) to the sample.

9. Incubate for 45 minutes at 4°C in the dark.

10. Add 500 μl 1X Permeabilization Wash Buffer. Centrifuge, remove the supernatant and vortex. repeat.

11. Resuspend it in staining buffer, and then perform flow cytometry analysis.

12. Once staining is complete, get the cells and record the percentage of ^{FOXP3+ positive cells.}

CTG determination method:

1. Thaw 1 vial aliquot of CTG reagent at room temperature and allow it to equilibrate to room temperature before use.

2. At room temperature, equilibrate the assay plate for approximately 20 minutes.

3. Collect the supernatant 120 μ L culture dish to another, for IL-10 ELISA analysis.

4. Add CTG agent 30 μ L / hole of the measuring cell culture dishes.

5. At room temperature, incubate on a culture plate shaker at 300 rpm for 15 minutes to avoid light exposure.

6. Read the luminescence signal on the Envision culture plate reader.

data analysis

The inhibition rate of the compound is calculated according to the following formula:% inhibition=100-100×(luminescence value-low luminescence value)/(high luminescence value-low luminescence value).

ELISA assay procedure:

˙Reagent preparation:

1. Dilute 5× Coating Buffer A to 1× with deionized water. For a culture plate, dilute 2.4 mL of 5× coating buffer in 9.6 mL of deionized water.

2. Dilute the captured antibody for preliminarily determined 1:200 in 1×coating buffer A. For a culture plate, 1 × coated on 11.94mL of Buffer A, 60 μ L diluted capture antibody.

3. Dilute 5× Assay Diluent A to 1× with PBS (pH 7.4). For 50 mL, dilute 10 mL of 5×Assay Diluent A in 40 mL PBS.

4. The lyophilized vial is under vacuum pressure. Reconstitute the freeze-dried standard with 0.2 mL of 1× Determination Diluent A. Let the reconstituted standard stand at room temperature for 15 minutes, and then mix gently to make a dilution.

5. Prior to use, assay diluent to 1 × A, at μ L stock solution prepared at a concentration of 1,000 250pg / mL of the highest standard (see Lot-Specific Cettificate of Analysis / ELISA MAX TM Deluxe Set Protocol).

In different test tubes, perform six two-fold serial dilutions of the 250pg/mL highest concentration standard with 1×Assay Diluent A. After dilution, the human IL-10 standard concentrations were 250pg/mL, 125pg/mL, 62.5pg/mL, 31.3pg/mL, 15.6pg/mL, 7.8pg/mL, and 3.9pg/mL, respectively. 1×Assay Diluent A as the zero standard (0pg/mL).

6. Dilute the biotinylated detection antibody for the preliminary determination at 1:200 in 1×Assay Diluent A. On a culture plate, 60 μ L of detection antibody diluted in diluent A 11.94mL 1 × assay.

7. Dilute avidin-HRP at 1:1000 in 1×Assay Diluent A. On a culture plate, 12 μ L of avidin A -HRP diluted in diluent 11.99mL 1 × assay.

8. TMB matrix solution is a mixture of matrix solution A and matrix solution B of equal volume. The two ingredients are mixed just before use. For a culture plate, in a clean container, mix 5.5 mL of matrix solution A with 5.5 mL of matrix solution B (the solution should be transparent and colorless).

9. Sample: Regarding the cell culture supernatant sample, the user may need to determine the dilution factor in the preliminary experiment. Serum or plasma samples should first be tested without any dilution. If dilution is required, the sample should be diluted in 1×Assay Diluent A before adding to the wells.

˙ELISA measurement:

1. The day before the ELISA, dilute the capture antibody in 1× Coating Buffer A as described in the reagent preparation. Add 100 μ L of this capture all of the 96 well plates provided by the antibody solution to this group. Seal the plate and incubate at 2°C and 8°C overnight (16 to 18 hours).

2. Before use, put all reagents at room temperature (RT). It is strongly recommended that all standards and samples be performed in two or three repetitions. Each test requires a standard curve.

3. Each well was washed with a buffer of at least 300 μ L culture plate was washed four times, and then via the plates were inverted on absorbent paper to firmly pat dry residue of buffer. All subsequent washings should be carried out in the same way.

4. For blocking non-specific binding and to reduce the background value, each well was added 200 μ L 1 × assay diluent A.

5. Seal the culture plate and incubate for 1 hour at room temperature on a culture plate shaker (for example, 500 rpm, 0.3 cm circular orbit). All subsequent shaking cultivations should be carried out in the same way.

6. While blocking the culture plate, prepare appropriate sample diluent (if necessary) and standards.

7. Wash the culture plate 4 times with washing buffer.

8. Add 100 μ L / well standard or sample product to the appropriate wells. If dilution is required, the sample should be diluted in 1×Assay Diluent A before adding to the wells.

9. Seal the culture plate and incubate with shaking at room temperature for 2 hours.

10. Wash the culture plate 4 times with washing buffer.

11. Add 100 μ L of diluted antibody was detected in each well, sealing plate culture and incubated shaking at room temperature for 1 hour.

12. Wash the culture plate 4 times with washing buffer.

13. Add 100 μ L of diluted avidin -HRP solution to each well, sealing plate culture and incubated shaking at room temperature for 30 minutes.

14. Wash the culture plate 5 times with washing buffer. For this final wash, immerse the wells in wash buffer for 30 seconds to 1 minute per wash. This will help minimize the background value.

15. Add 100 μ L of freshly mixed TMB substrate solution, and incubated in the dark for 30 minutes. The color of the positive hole should change to blue. There is no need to seal the culture plate during this step.

16. The reaction by the addition of 100 μ L of stop solution to each well to terminate. The positive hole should change from blue to yellow.

17. Read the absorbance at 450nm within 15 minutes. If the reader can read the absorbance at 570nm, the absorbance at 570nm can be subtracted from the absorbance at 450nm.

Using the above assay, the inhibitory effects of several representative compounds of the present invention (and the comparative compounds in several cases) on Treg cells are summarized in the following tables. Since the determinations performed in different batches may not be directly comparable with each other, only the compounds analyzed in the same batch can be directly compared in the same table. It should be noted that the compounds in the comparison group are partial inhibitors of Treg inhibition, because the maximum inhibition that can be achieved at the highest concentration is about 55 to 65%.

Similarly, the inhibitory effects of several representative compounds of the present invention (and the comparative compounds in several cases) on Th17 cells are summarized in the following table.

Example 4. Pharmacokinetic (PK) data in mice

The pharmacokinetic data of the representative compound of the present invention and the comparison group compound were obtained from CD1 male mice. Specifically, the CD1 male mice of each group were fed orally with a low dose of 5 mg/kg of the compound or a high dose of 30 mg/kg of the same compound orally. Obtain PK data for various compounds of the present invention and the comparison group, including AUC _last (hr*ng/mL) and bioavailability (F%). Based on the high-dose and low-dose AUC of the same compound, calculate the multiple of increase in AUC.

B. Synthesis Example

Device Description

The NMR spectroscopy system uses a Varian Mercury spectrometer, operating at 400 MHz ( ¹ H), 376 MHz ( ¹⁹ F), or 75 MHz ( ¹³ C). The solvent used for the sample is specified in the experimental procedure of each compound. The chemical shift is expressed in parts per million (ppm, δ unit). The coupling constant is in Hertz (Hz). Split pattern depicts obvious multiplicity, and named s (single peak), d (double peak), t (triplet), q (quartet), quint (quintet), m (multiple peak), br (Broad peak).

LCMS uses the following system: Agilent 6120 (binary gradient module pump), XBridge analytical column C ₁₈ , 5 μ m, 4.6×50 mm, 25° C., 5 μ L injection volume, 2 mL/min, acetonitrile in 0.1 The gradient in %ammonium acetate aqueous solution is carried out according to the following timing:

Experimental procedure : Unless otherwise specified, all reactions were carried out under a dry nitrogen atmosphere. The TLC panel is exposed to ultraviolet light. Flash chromatography refers to the use of glass column on silica gel (40 to 60μm) to perform column chromatography. Alternatively, the Biotage SP1 or Biotage Isolera system for uv detection at 220 or 254 nm and the use of Biotage normal phase or reverse phase silica gel cartridges for automated chromatography can be used. Further details can be found in the relevant experimental procedures.

Process 1

Synthesis of compound of formula I

酸9於二苯基磷醯基疊氮化物(DPPA)存在下進行Curtis重排，並以吡啶胺10處理，得到式I化合物。 The compound of formula I can be prepared as described in Scheme 1 as an imidazole intermediate. In the presence of a base such as sodium hydride, the imidazole derivative 1, such as (amine oxide) diphenylphosphine oxide, is reacted in a solvent such as DMF to obtain an aminated imidazole derivative 2 . Compound 2 can be N-protected by reacting with dicarbonate di(tertiary butyl) to give compound 3 . Compound 3 can be brominated or chlorinated with a brominating or chlorinating agent, such as bromine or N-bromosuccinimide, in a solvent such as DMF to obtain compound 4 as the main product. Compound 4 can be reacted with ethyl acetate in a solvent such as THF in the presence of a base such as potassium tertiary butoxide to obtain compound 5 . At room temperature, in a solvent such as DCM, the compound 5 is reacted with 1,1-dimethoxy-N,N-dimethylmethylamine (DMF-DMA) to obtain the cyclized product 6 . Compound 6 is reacted with a chlorinating agent, such as phosphorous oxy chloride/bromide (phosphorus oxy chloride/bromide), to obtain compound 7. Compound 7 can be reacted with boric acid or boric acid ester of _{formula R 1} B(OH) ₂ under Suzuki coupling conditions to obtain compound 8 . Compound 8 can be hydrolyzed in a protic solvent such as methanol or ethanol with a hydrolyzing agent such as lithium hydroxide or sodium hydroxide to obtain acid derivatives 9 . Imidazolium

Curtis rearrangement of acid 9 in the presence of diphenylphosphoryl azide (DPPA) and treatment with pyridylamine 10 yields the compound of formula I.

Process 2

Synthesis of compound of formula II

醚類似物11，將其以LiOH處理，得到酸12。咪唑嗒

酸9於二苯基磷醯基疊氮化物(DPPA)存在下進行Curtis重排，以吡啶胺10處理，得到式II化合物。 After one or more solvents such as DMF, DMA, THF, toluene or a mixture thereof, use an appropriate base such as sodium hydride, lithium hexamethyldisilane, cesium carbonate, potassium carbonate, and alkyl halides (such as methyl iodide, Ethane, propyl bromide) to alkylate compound 6 to obtain imidazolium

Ether analog 11 is treated with LiOH to obtain acid 12 . Imidazole

Curtis rearrangement of acid 9 in the presence of diphenylphosphoryl azide (DPPA) and treatment with pyridylamine 10 gives the compound of formula II.

Process 3

Synthesis of compound of formula III

烷、或甲苯中，以一級胺或二級胺R₅R₆NH進行溴化物7之Buchwald-Hartwig胺化反應，最後得到式III之化合物。 The compounds of formula I and II can be prepared following the method described in Scheme 3. For suitable Pd catalysts such as Pd ₂ dba ₃ , Pd(dppf)Cl ₂ , Pd(OAc) ₂ , suitable ligands such as BINAP, xantphos, triphenylphosphine and suitable bases such as t-BuONa or t-BuOK are present Under the solvent such as tetrahydrofuran, two

In alkane or toluene, the Buchwald-Hartwig amination reaction of bromide 7 is carried out with the primary amine or the secondary amine R ₅ R ₆ NH, and finally the compound of formula III is obtained.

Process 4

Synthesis of compound of formula IV

The compound of formula IV can be prepared following the method described in Scheme 4. Under Pd/C and hydrogen, compound 7 was hydrogenated to obtain compound 15 , which was hydrolyzed with lithium hydroxide, followed by Curtis rearrangement, and quenched with aminopyridine 10 or derivatives, to finally obtain compound of formula IV .

Process 5

Synthesis of compound of formula V

酯7製備。鹼性條件(例如K₂CO₃、LiOH、NaOH等)下，於溶劑如THF、水、甲醇或其混合物中，酯7之水解得到對應之碳環酸(carbocylic acid)17，其於DPPA與三級胺鹼存在下進行Curtis重排，並捕捉異氰酸酯衍生物，得到所需產物10。隨後溴化物18與有機錫化合物 Stille之偶合得到對應之炔烴化合物19，將其以HCl水溶液處理，得到酮20；以硼氫化鈉還原成對應之醇21後，用亞硫醯氯處理，然後以不同醇猝滅，得到醚22，令其與胺甲酸酯(carbomate)23反應，得到脲化合物V。 The compound of formula V can be as described in Scheme 5 with imidazolium

Ester 7 preparation. Under alkaline conditions (such as K ₂ CO ₃ , LiOH, NaOH, etc.), in a solvent such as THF, water, methanol or a mixture thereof, the ester 7 is hydrolyzed to obtain the corresponding carbocylic acid 17 , which is compatible with DPPA Curtis rearrangement is performed in the presence of a tertiary amine base and the isocyanate derivative is captured to obtain the desired product 10. Then the bromide 18 is coupled with the organotin compound Stille to obtain the corresponding alkyne compound 19 , which is treated with aqueous HCl to obtain the ketone 20 ; after reducing it to the corresponding alcohol 21 with sodium borohydride, it is treated with sulfite chloride, and then It is quenched with different alcohols to obtain ether 22 , which is reacted with carbomate 23 to obtain urea compound V.

Intermediate products 6c and 7c

step 1

At 0°C, NaH (3.55g, 92.76mmol, 60% purity) was added to 1H-imidazole-2-carboxylic acid ethyl ester (10g, 71.36mmol) in THF (200mL) solution, and the resulting solution was warmed to room temperature , Stirred for 2 hours, then cooled to 0°C, and added dropwise a solution of O-diphenylphosphoryl hydroxylamine (23.30 g, 99.90 mmol) in THF (200 mL) to the mixture. After 20 minutes, the ice bath was removed, and the resulting mixture was stirred at room temperature for 12 hours. TLC showed the formation of new spots; the reaction was quenched with water until it became a clear solution and concentrated in vacuo to dryness. After stirring for 30 minutes, the crude product was suspended in ethyl acetate (700 mL) and filtered. The filtrate was concentrated and purified by flash column chromatography (DCM: methanol 30:1) to obtain ethyl 1-aminoimidazole-2-carboxylate (10.32 g, 66.51 mmol, 93.21% yield) as a colorless oil. LC-MS: m/z 156 [M+H] ⁺ .

Step 2

To the stirred solution of ethyl 1-aminoimidazole-2-carboxylate (10g, 64.45mmol) in DMF (60mL), add DMAP (1.18g, 9.67mmol) and bis(tertiary butyl) dicarbonate (18.29g) , 83.79 mmol, 19.23 mL). The reaction mixture was stirred at 85°C for 4 hours. After the reaction was complete, the mixture was poured into water and extracted with EtOAc (2×600 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (petroleum ether: ethyl acetate = 5:1) to obtain ethyl 1-(tertiary butoxycarbonylamino)imidazole-2-carboxylate (13.2g, 51.71mmol, 80.23% yield), as a yellow solid. LC-MS: m/z 256[M+H]+.

Step 3

In a suspension of 1-(tertiary butoxycarbonylamino)imidazole-2-carboxylic acid ethyl ester (3.5g, 13.71mmol) in DMF (15mL), slowly add 1-chloropyrrole in DMF (5mL) Pyridine-2,5-dione (1.83 g, 13.71 mmol, 1.11 mL). The reaction mixture was stirred at room temperature for 6 hours. After the reaction was complete, saturated sodium bicarbonate was added to the reaction and extracted with EtOAc (2×150 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=10:1) to obtain 1-(tertiary butoxycarbonylamino)-4-chloro-imidazole-2-carboxylic acid ethyl ester (1.9 g, 6.56 mmol, 47.83% yield), as a colorless oil. LC-MS: m/z 290 [M+H] ⁺ .

Step 4

At 0° C., in a THF (10 mL) solution of 1-(tertiary butoxycarbonylamino)-4-chloro-imidazole-2-carboxylate (3.7 g, 12.77 mmol) in THF (10 mL), 2-methyl was added dropwise Potassium propane-2-oleate (1M, 38.31 mL) for 5 minutes. Anhydrous ethyl acetate (2.81 g, 31.93 mmol, 3.12 mL) was added to the mixture, and the mixture was stirred at 0°C for 15 minutes. The ice bath was removed, and the resulting mixture was stirred at room temperature for 2 hours. TLC showed the formation of spots, the reaction was cooled to 0°C, treated with 1.0 N HCl via dropwise addition until the suspension was completely dissolved, and then extracted with EtOAc (2×40 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain (Z)-3-[1-(tertiary butoxycarbonylamino)-4-chloro-imidazole-2 -Yl]-3-hydroxy-prop-2-enoic acid ethyl ester (4.24 g, crude product) as a yellow solid. LC-MS: m/z 332[M+H] ⁺ .

Step 5

-7-羧酸乙酯(2.4g，9.93mmol，77.72%產率)，為白色固體。LC-MS：m/z 242[M+H]⁺。 To (Z)-3-[1-(tertiary butoxycarbonylamino)-4-chloro-imidazol-2-yl]-3-hydroxy-prop-2-enoic acid ethyl ester (4.24g, 12.78mmol) To the DCM (30 mL) solution, 1,1-dimethoxy-N,N-dimethyl-methylamine (12.18 g, 102.25 mmol, 13.69 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was concentrated to obtain a crude product, which was recrystallized with ethyl acetate to obtain 2-chloro-8-hydroxy-imidazo[1,2-b]

Ethyl -7-carboxylate (2.4 g, 9.93 mmol, 77.72% yield), a white solid. LC-MS: m/z 242 [M+H] ⁺ .

Step 6

-7-羧酸乙酯(6g，24.83mmol)之1,4-二

烷(20mL)攪拌溶液中，添加磷醯溴(17.80g，62.08mmol，6.31mL)，於110℃攪拌反應混合物2小時，反應完全後，冷卻反應混合物至室溫，將其傾入水中，以EtOAc(2×150mL)萃取。合併之有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物利用快速管柱層析法(石油醚/乙酸乙酯= 7：1)純化，得到8-溴-2-氯-咪唑并[1,2-b]嗒

-7-羧酸乙酯(3.3g，10.84mmol，43.64%產率)，為黃色固體。LC-MS：m/z 304[M+H]⁺。 At room temperature, in 2-chloro-8-hydroxy-imidazo[1,2-b]

-7-carboxylic acid ethyl ester (6g, 24.83mmol) of 1,4-bis

Add phosphonium bromide (17.80g, 62.08mmol, 6.31mL) to the stirring solution of alkane (20mL), and stir the reaction mixture at 110°C for 2 hours. After the reaction is complete, cool the reaction mixture to room temperature and pour it into water. Extract with EtOAc (2×150 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate = 7:1) to obtain 8-bromo-2-chloro-imidazo[1,2-b]

Ethyl -7-carboxylate (3.3 g, 10.84 mmol, 43.64% yield) as a yellow solid. LC-MS: m/z 304 [M+H] ⁺ .

-7-基)脲之合成 Example 1. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-cyclopropylimidazo [1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(700.0mg，2.31mmol)之甲苯(42mL)溶液中，添加環丙基硼酸(397.4mg，4.62mmol)、三環己膦(646.8mg，2.31mmol)與磷酸鉀(1.22g，5.775mmol)。將懸浮液脫氣並與N₂交換兩次。添加乙酸鈀(II)(129.4mg，0.5775mmol)至該混合物中。加熱反應混合物至100℃ 12小時。耗盡起始原料並以LC-MS檢測到所需質量。減壓濃縮混合物，所得殘留物以快速管柱層析法Z(以於石油中1%至15%之乙酸乙酯洗提)純化，得到2-氯-8-環丙基咪唑并[1,2-b]嗒

-7-羧酸乙酯(330.0mg，50.3%產率)，為黃色固體。LC-MS：m/z 266[M+H]⁺。 On 8-bromo-2-chloroimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 7c (700.0mg, 2.31mmol) in toluene (42mL), add cyclopropylboronic acid (397.4mg, 4.62mmol), tricyclohexylphosphine (646.8mg, 2.31mmol) and potassium phosphate (1.22g, 5.775mmol). The suspension was degassed and _{exchanged with N 2} twice. Palladium(II) acetate (129.4 mg, 0.5775 mmol) was added to the mixture. The reaction mixture was heated to 100°C for 12 hours. The starting material was consumed and the required quality was detected by LC-MS. The mixture was concentrated under reduced pressure, and the resulting residue was purified by flash column chromatography Z (eluted with 1% to 15% ethyl acetate in petroleum) to obtain 2-chloro-8-cyclopropylimidazo[1, 2-b]

Ethyl -7-carboxylate (330.0 mg, 50.3% yield), as a yellow solid. LC-MS: m/z 266 [M+H] ⁺ .

Step 2

-7-羧酸乙酯(352mg，1.328mmol)之THF(8mL)溶液中，添加氫氧化鋰一水合物(111.6mg，2.657mmol)之水(2.5mL)溶液。於30℃攪拌混合物4小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮去除溶劑；殘留物以HCl(1M)酸化至pH=3~4。以EtOAc萃取混合物，乾燥有機相，濃縮，得到2-氯-8-環丙基咪唑并[1,2-b]嗒

-7-羧酸(310mg，98.5%產率)，為黃色固體。LC-MS：m/z 238[M+H]⁺。 On 2-chloro-8-cyclopropylimidazo[1,2-b]

To a solution of ethyl -7-carboxylate (352 mg, 1.328 mmol) in THF (8 mL), a solution of lithium hydroxide monohydrate (111.6 mg, 2.656 mmol) in water (2.5 mL) was added. The mixture was stirred at 30°C for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. Concentrate to remove the solvent; the residue is acidified with HCl (1M) to pH=3~4. The mixture was extracted with EtOAc, the organic phase was dried, and concentrated to obtain 2-chloro-8-cyclopropylimidazo[1,2-b]

-7-carboxylic acid (310 mg, 98.5% yield), as a yellow solid. LC-MS: m/z 238 [M+H] ⁺ .

Step 3

-7-羧酸(150mg，0.6329mmol)之1,4-二

烷(10mL)溶液中，添加疊氮膦酸二苯酯(diphenylphosphonic azide)(209mg，0.7595mmol)與三乙胺(320.2mg，3.164mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(185.1mg，0.9494mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。有機相以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-環丙基咪唑并[1,2-b]嗒

-7-基)脲(64mg)，為白色固體。LC-MS：m/z 430[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.78(s,1 H),8.99(s,1 H),8.72(s,1 H),8.58(d,J=2.4Hz,1 H),8.48(d,J=2.4Hz,1 H),8.34(s,1 H),8.16(s,2 H),2.17-2.14(m,1 H),1.73-1.71(m,2 H),1.16-1.14(m,2 H)。 On 2-chloro-8-cyclopropylimidazo[1,2-b]

-7-carboxylic acid (150mg, 0.6329mmol) of 1,4-bis

To the alkane (10 mL) solution, diphenylphosphonic azide (209 mg, 0.7595 mmol) and triethylamine (320.2 mg, 3.164 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (185.1 mg, 0.9494 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic phase was _{dried with Na 2} SO ₄ and concentrated. The residue obtained was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(5-chloro-6- (2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-cyclopropylimidazo[1,2-b]ta

-7-yl)urea (64 mg) as a white solid. LC-MS: m/z 430 [M+H] ⁺ . ¹ H NMR (DMSO- d6 ,400MHz): δ 9.78 (s, 1 H), 8.99 (s, 1 H), 8.72 (s, 1 H), 8.58 (d, J = 2.4 Hz, 1 H), 8.48 (d, J = 2.4Hz, 1 H), 8.34 (s, 1 H), 8.16 (s, 2 H), 2.17-2.14 (m, 1 H), 1.73-1.71 (m, 2 H), 1.16 1.14 (m, 2 H).

-7-基)脲之合成 Example 2. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-isopropylimidazo [1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯(250mg，820.94μmol)、2-異丙烯基-4,4,5,5-四甲基-1,3,2-二

硼烷7c(179.3mg，1.07mmol)之甲苯(3mL)與水(0.3mL)溶液中，添加磷酸鉀(522.8mg，2.46mmol)與氯化雙(三苯膦)鈀(II)(57.6mg，82.09μmol)。N₂下，於100℃攪拌混合物24小時。LC-MS顯示殘留一些起始原料，再添加氯化雙(三苯膦)鈀(II)(57.6mg，82.09μmol)。N₂下，於100℃再攪拌混合物24小時。冷卻至室溫後，過濾反應混合物濃縮濾液，得到殘留物，利用快速管柱層析法(以PE/EA=10/1洗提)將其純化，得到為黃色固體之2-氯-8-異丙烯基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(57mg，26%產率)及回收100mg 8-溴-2-氯-咪唑并[1,2-b]嗒

-7-羧酸乙酯。LC-MS：m/z 266[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

Ethyl -7-carboxylate (250 mg, 820.94 μ mol), 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-di

To a solution of borane 7c (179.3mg, 1.07mmol) in toluene (3mL) and water (0.3mL), add potassium phosphate (522.8mg, 2.46mmol) and bis(triphenylphosphine) palladium(II) chloride (57.6mg) , 82.09 μ mol). The mixture was stirred at 100°C for 24 hours under _{N 2.} LC-MS showed some residual starting material, then add bis (triphenylphosphine) palladium (II) (57.6mg, 82.09 μ mol). Under N _2, the mixture was stirred at 100 deg.] C for 24 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated to obtain the residue, which was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8- as a yellow solid Isopropenyl-imidazo[1,2-b]

Ethyl -7-carboxylate (57 mg, 26% yield) and recovery 100 mg 8-bromo-2-chloro-imidazo[1,2-b]

-7-Ethyl carboxylate. LC-MS: m/z 266 [M+H] ⁺ .

Step 2

-7-羧酸乙酯(68mg，255.93μmol)之甲醇(5mL)溶液中，添加PtO₂(15mg，255.93μmol)。於室溫，攪拌反應混合物，於H₂下隔夜，然後過濾，真空濃縮。粗產物利用快速管柱層析法(以PE/EA =10/1洗提)純化，得到2-氯-8-異丙基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(32mg，47%產率)，為清油。LC-MS：m/z 268[M+H]⁺。 On 2-chloro-8-isopropenyl-imidazo[1,2-b]ta

To a methanol (5 mL) solution of ethyl -7-carboxylate (68 mg, 255.93 μ _{mol), PtO 2} (15 mg, 255.93 μ mol) was added. At room temperature, the reaction mixture was stirred _{under H 2} overnight, then filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (eluted with PE/EA = 10/1) to obtain 2-chloro-8-isopropyl-imidazo[1,2-b]

Ethyl -7-carboxylate (32mg, 47% yield), clear oil. LC-MS: m/z 268 [M+H] ⁺ .

Step 3

-7-羧酸乙酯(32mg，119.53μmol)之THF(2mL)溶液中，添加氫氧化鋰一水合物(20.1mg，478.13μmol)之水(0.4mL)溶液。於室溫攪拌混合物5小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取混合物，乾燥有機相，濃縮，得到為白色固體之2-氯-8-異丙基-咪唑并[1,2-b]嗒

-7-羧酸(28.6mg，定量)，直接用於下一步驟。LC-MS：m/z 240[M+H]⁺。 To 2-chloro-8-isopropyl-imidazo[1,2-b]

7-carboxylate (32mg, 119.53 μ mol) of THF (2mL) was added lithium hydroxide monohydrate (20.1mg, 478.13 μ mol) of water (0.4 mL of) solution. The mixture was stirred at room temperature for 5 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc, the organic phase was dried, and concentrated to obtain 2-chloro-8-isopropyl-imidazo[1,2-b]a as a white solid

-7-carboxylic acid (28.6 mg, quantitative), used directly in the next step. LC-MS: m/z 240 [M+H] ⁺ .

Step 4

烷(6mL)溶液中，添加疊氮膦酸二苯酯(37.1mg，143.20μmol)與三乙胺(60.4mg，596.68μmol)。所得溶液於室溫攪拌30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(28.0mg，143.20μmol)，於100℃攪拌混合物2小時。去除溶劑，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)與製備型HPLC純化，得到1-(2-氯-7-異丙基-吡唑并[1,5-a]嘧啶-6-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(8mg，18.51μmol，15.51%產率)，為白色固體。LC-MS：m/z 432.0[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.72(s,1H),8.91(s,1H),8.71(s,1H),8.57(d,J=2.4Hz,1H),8.47(d,J=2.4Hz,1H),8.39(s,1H),8.16(s,2H),3.51-3.44(m,1H),1.48(d,J=6.8Hz,6H)。 In 2-chloro-7-isopropyl-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid (28.6mg, 119.34 μ mol) of 1,4-di

To the alkane (6 mL) solution, add diphenyl azide phosphonate (37.1 mg, 143.20 μ mol) and triethylamine (60.4 mg, 596.68 μ mol). The resulting solution was stirred at room temperature for 30 minutes and then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (28.0mg, 143.20 μ mol), the mixture was stirred at 100 deg.] C for 2 hours. The solvent was removed, and the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) and preparative HPLC to obtain 1-(2-chloro-7-isopropyl-pyrazolo[1, 5-a] pyrimidin-6-yl) -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (8mg, 18.51 μ mol, 15.51 % yield) as a white solid. LC-MS: m/z 432.0 [M+H] ⁺ . ¹ H NMR(DMSO- d6 ,400MHz): δ 9.72(s,1H),8.91(s,1H),8.71(s,1H),8.57(d, J =2.4Hz,1H), 8.47(d, J =2.4Hz, 1H), 8.39 (s, 1H), 8.16 (s, 2H), 3.51-3.44 (m, 1H), 1.48 (d, J = 6.8 Hz, 6H).

-7-基)脲之合成 Example 3. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-ethylimidazo[ 1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(600mg，1.97mmol)、4,4,5,5-四甲基-2-乙烯基-1,3,2-二

硼烷(910mg，5.91mmol)與三環己膦(552mg，1.97mmol)之1,4-二

烷(20mL)溶液中，添加磷酸鉀(1.25g，5.91mmol)與乙酸鈀(II)(110mg，492.57μmol)。N₂下，於100℃攪拌該懸浮液2小時。冷卻至室溫後，過濾反應混合物，濃縮濾液，得到殘留物，利用快速管柱層析法(以PE/EA=10/1洗提)將其純化，得到2-氯-8-乙烯基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(230mg，46%產率)，為黃色固體。LC-MS：m/z 252[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

Ethyl -7-carboxylate 7c (600mg, 1.97mmol), 4,4,5,5-tetramethyl-2-vinyl-1,3,2-di

Borane (910mg, 5.91mmol) and tricyclohexylphosphine (552mg, 1.97mmol) 1,4-bis

Dioxane (20mL) was added potassium phosphate (1.25g, 5.91mmol) and palladium acetate (II) (110mg, 492.57 μ mol). Under N _2, was stirred at 100 deg.] C the suspension for 2 hours. After cooling to room temperature, the reaction mixture was filtered and the filtrate was concentrated to obtain the residue, which was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-vinyl- Imidazo[1,2-b]

Ethyl -7-carboxylate (230 mg, 46% yield), as a yellow solid. LC-MS: m/z 252 [M+H] ⁺ .

Step 2

-7-羧酸乙酯8d(230mg，913.90μmol)之甲醇(5mL)溶液中，添加PtO₂(41.5mg，182.78μmol)。H₂下，於室溫攪拌反應混合物3小時，然後過濾，真空濃縮。粗產物利用快速管柱層析法(以PE/EA=10/1洗提)純化，得到2-氯-8-乙基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(170mg，73%產率)，為清油。LC-MS：m/z 254[M+H]⁺。 On 2-chloro-8-vinyl-imidazo[1,2-b]ta

To a methanol (5 mL) solution of ethyl -7-carboxylate 8d (230 mg, 913.90 μ _{mol), PtO 2} (41.5 mg, 182.78 μ mol) was added. Under H _2, the reaction mixture was stirred at room temperature for 3 hours, then filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-ethyl-imidazo[1,2-b]

Ethyl -7-carboxylate (170 mg, 73% yield), a clear oil. LC-MS: m/z 254 [M+H] ⁺ .

Step 3

-7-羧酸乙酯8e(170mg，670.12μmol)之THF(2mL)與乙醇(1mL)溶液中，添加氫氧化鋰一水合物(112.5mg，2.68mmol)之水溶液(1mL)。於室溫攪拌混合物4小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取該混合物，乾燥有機相，濃縮，得到2-氯-8-乙基-咪唑并[1,2-b]嗒

-7-羧酸(150mg，99%產率)，為白色固體，直接用於下一步驟。LC-MS：m/z 226[M+H]⁺。 On 2-chloro-8-ethyl-imidazo[1,2-b]

7-carboxylic acid ethyl ester 8e (170mg, 670.12 μ mol) of THF (2mL) and ethanol (1 mL) was added lithium hydroxide monohydrate (112.5mg, 2.68mmol) of an aqueous solution (1mL). The mixture was stirred at room temperature for 4 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc, the organic phase was dried and concentrated to give 2-chloro-8-ethyl-imidazo[1,2-b]

-7-carboxylic acid (150 mg, 99% yield), a white solid, was used directly in the next step. LC-MS: m/z 226 [M+H] ⁺ .

Step 4

烷(6mL)溶液中，添加疊氮膦酸二苯酯(137.8mg，531.84μmol)與三乙胺(224.2mg，2.22mmol)。所得溶液於室溫攪拌30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(130.0mg，664.80μmol)，於100℃攪拌混合物2小時。去除溶劑，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)與製備型HPLC純化，得到1-(2-氯-7-乙基-吡唑并[1,5-a]嘧啶-6-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(68mg，37%產率)。LC-MS：m/z 418.0[M+H]^{+ 1}H NMR(DMSO-d6,400MHz)：δ 9.78(brs,1H),8.93(brs,1H),8.90(s,1H),8.58(d,J=2.4Hz,1H),8.48(d,J=2.4Hz,1H),8.38(s,1H),8.16(s,2H),2.95(q,J=7.2Hz,2H),1.26(t,J=7.6Hz,3H)。 In 2-chloro-7-ethyl-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid 9c (100mg, 443.20 μ mol) of 1,4-di

Dioxane (6mL) was added azide diphenyl phosphonate (137.8mg, 531.84 μ mol) and triethylamine (224.2mg, 2.22mmol). The resulting solution was stirred at room temperature for 30 minutes and then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (130.0mg, 664.80 μ mol), the mixture was stirred at 100 deg.] C for 2 hours. The solvent was removed, and the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) and preparative HPLC to obtain 1-(2-chloro-7-ethyl-pyrazolo[1,5 -a]pyrimidin-6-yl)-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (68 mg, 37% yield). LC-MS: m/z 418.0[M+H] ^{+ 1} H NMR (DMSO- d6 ,400MHz): δ 9.78(brs,1H), 8.93(brs,1H), 8.90(s,1H), 8.58(d , J =2.4Hz,1H),8.48(d, J =2.4Hz,1H),8.38(s,1H),8.16(s,2H),2.95(q, J =7.2Hz,2H),1.26(t , J =7.6Hz,3H).

-7-基)-3-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)脲之合成 Example 4. 1-(8-Bromo-2-chloroimidazo[1,2-b]

Synthesis of -7-yl)-3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)urea

step 1

-7-羧酸乙酯9d(114mg，0.3762mmol)之THF(4mL)溶液中，添加氫氧化鋰一水合物(23.7mg，0.5644mmol)之水(0.5mL)溶液。於30℃攪拌混合物4小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮所得混合物以移除溶劑，殘留物以HCl(1M)酸化至pH=3~4。以EtOAc萃取混合物，乾燥有機相，濃縮，得到8-溴-2-氯咪唑并[1,2-b]嗒

-7-羧酸(96mg，92.8%)，為黃色固體。LC-MS：m/z 276[M+H]⁺。 On 8-bromo-2-chloroimidazo[1,2-b]

To the THF (4 mL) solution of ethyl-7-carboxylate 9d (114 mg, 0.3762 mmol), was added lithium hydroxide monohydrate (23.7 mg, 0.5644 mmol) in water (0.5 mL). The mixture was stirred at 30°C for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. The resulting mixture was concentrated to remove the solvent, and the residue was acidified with HCl (1M) to pH=3~4. The mixture was extracted with EtOAc, the organic phase was dried and concentrated to give 8-bromo-2-chloroimidazo[1,2-b]

-7-carboxylic acid (96 mg, 92.8%), as a yellow solid. LC-MS: m/z 276 [M+H] ⁺ .

Step 2

-7-羧酸(96mg，0.3491mmol)之1,4-二

烷(6mL)溶液中，添加疊氮膦酸二苯酯(115.3mg，0.4189mmol)與三乙胺(176.6mg，1.745mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(102.1mg，0.5236mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。該有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(8-溴-2-氯咪唑并[1,2-b]嗒

-7-基)-3-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)脲(10.2mg)，為白色固體。LC-MS：m/z 468[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 10.19(s,1 H),9.13(s,1 H),9.06(s,1 H),8.57(d,J=1.6Hz,1 H),8.54(s,1 H),8.49(d,J=2.0Hz,1 H),8.17(s,2 H)。 On 8-bromo-2-chloroimidazo[1,2-b]

-7-carboxylic acid (96mg, 0.3491mmol) of 1,4-bis

To the alkane (6 mL) solution, add diphenyl azide phosphonate (115.3 mg, 0.4189 mmol) and triethylamine (176.6 mg, 1.745 mmol). The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (102.1 mg, 0.5236 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The residue obtained was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(8-bromo-2- Chlorimidazo[1,2-b]ta

-7-yl)-3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)urea (10.2 mg) as a white solid. LC-MS: m/z 468 [M+H] ⁺ . ¹ H NMR (DMSO- d6 ,400MHz): δ 10.19 (s, 1 H), 9.13 (s, 1 H), 9.06 (s, 1 H), 8.57 (d, J = 1.6 Hz, 1 H), 8.54 (s, 1 H), 8.49 (d, J = 2.0 Hz, 1 H), 8.17 (s, 2 H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 5. 1-(2-Chloro-8-cyclopropylimidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

step 1

-7-羧酸9a(50mg，0.211mmol)之1,4-二

烷(2mL)溶液中，添加疊氮膦酸二苯酯(69.7mg，0.2532mmol)與三乙胺(106.8mg，1.055mmol)。於室溫攪拌所得混合物30分鐘，然後添加2-(三氟甲基)吡啶-4-胺10b(51.3mg，0.317mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。該有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(2-氯-8-環丙基咪唑并[1,2-b]嗒

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲(5.2mg)，為白色固體。LC-MS：m/z 397[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.89(s,1 H),8.95(s,1 H),8.68(s,1 H),8.55(d,J=5.6Hz,1 H),8.34(s,1 H),8.07(d,J=1.6Hz,1 H),7.63-7.62(m,1 H),2.16-2.12(m,1 H),1.71-1.69(m,2 H),1.14-1.12(m,2 H)。 On 2-chloro-8-cyclopropylimidazo[1,2-b]

-7-carboxylic acid 9a (50mg, 0.211mmol) of 1,4-bis

To the alkane (2 mL) solution, diphenyl azide phosphonate (69.7 mg, 0.2532 mmol) and triethylamine (106.8 mg, 1.055 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, then 2-(trifluoromethyl)pyridine-4-amine 10b (51.3 mg, 0.317 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The residue obtained was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(2-chloro-8- Cyclopropylimidazo[1,2-b]

-7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea (5.2 mg) as a white solid. LC-MS: m/z 397 [M+H] ⁺ . ¹ H NMR(DMSO- d6 ,400MHz): δ 9.89(s,1 H),8.95(s,1 H),8.68(s,1 H),8.55(d, J =5.6Hz,1 H), 8.34 (s, 1 H), 8.07 (d, J = 1.6 Hz, 1 H), 7.63-7.62 (m, 1 H), 2.16-2.12 (m, 1 H), 1.71-1.69 (m, 2 H), 1.14-1.12 (m, 2 H).

-7-基)-3-(6-(三氟甲基)吡啶-3-基)脲之合成 Example 6. 1-(2-Chloro-8-cyclopropylimidazo[1,2-b]

Synthesis of -7-yl)-3-(6-(trifluoromethyl)pyridin-3-yl)urea

step 1

-7-羧酸9a(50mg，0.211mmol)之1,4-二

烷(3mL)溶液中，添加疊氮膦酸二苯酯(69.7mg，0.2532mmol)與三乙胺(106.8mg，1.055mmol)。於室溫攪拌所得混合物30分鐘，然後添加6-(三氟甲基)吡啶-3-胺10c(51.3mg，0.3165mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。有機相以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至100%之乙酸乙酯洗提)純化，得到1-(2-氯-8-環丙基咪唑并[1,2-b]嗒

-7-基)-3-(6-(三氟甲基)吡啶-3-基)脲(12.7mg)，為白色固體。LC-MS：m/z 397[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.69(s,1 H),8.88(s,1 H),8.76(d,J=2.0Hz,1 H),8.72(s,1 H),8.33(s,1 H),8.26-8.23(m,1 H),7.84(d,J=8.8Hz,1 H),2.16-2.12(m,1 H),1.72-1.68(m,2 H),1.15-1.11(m,2 H)。 On 2-chloro-8-cyclopropylimidazo[1,2-b]

-7-carboxylic acid 9a (50mg, 0.211mmol) of 1,4-bis

To the alkane (3 mL) solution, diphenyl azide phosphonate (69.7 mg, 0.2532 mmol) and triethylamine (106.8 mg, 1.055 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, then 6-(trifluoromethyl)pyridin-3-amine 10c (51.3 mg, 0.3165 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic phase was _{dried with Na 2} SO ₄ and concentrated. The resulting residue was purified by flash column chromatography (eluted with 10% to 100% ethyl acetate in petroleum) to obtain 1-(2-chloro-8- Cyclopropylimidazo[1,2-b]

-7-yl)-3-(6-(trifluoromethyl)pyridin-3-yl)urea (12.7 mg) as a white solid. LC-MS: m/z 397 [M+H] ⁺ . ¹ H NMR(DMSO- d6 ,400MHz): δ 9.69(s,1 H), 8.88(s,1 H), 8.76(d, J =2.0Hz,1 H), 8.72(s,1 H), 8.33 (s, 1 H), 8.26-8.23 (m, 1 H), 7.84 (d, J = 8.8Hz, 1 H), 2.16-2.12 (m, 1 H), 1.72-1.68 (m, 2 H), 1.15-1.11 (m, 2 H).

-7-基)-3-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)脲之合成 Example 7. 1-(2-Bromo-8-cyclopropylimidazo[1,2-b]

Synthesis of -7-yl)-3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)urea

step 1

In the suspension of 1-(tertiary butoxycarbonylamino)imidazole-2-carboxylic acid ethyl ester 3 (3.57g, 13.99mmol) in DMF (30mL), slowly add 1-bromopyrrolidine-2,5- A solution of diketone (2.49 g, 13.99 mmol, 1.19 mL) in DMF (10 mL). The reaction mixture was stirred at room temperature for 12 hours. After the reaction was judged to be complete by TLC, saturated sodium bicarbonate was added to the reaction mixture and extracted with EtOAc (2×150 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate, 10:1) to obtain ethyl 4-bromo-1-(tertiary butoxycarbonylamino)imidazole-2-carboxylate (2.5g , 7.48mmol, 53.49% yield), as a colorless oil. LC-MS: m/z 334.1 [M+H] ⁺ .

Step 2

At 0°C, in 4-bromo-1-(tertiary butoxycarbonylamino)imidazole-2-carboxylic acid ethyl ester 4d (2.5g, 7.48mmol) in THF (4mL) solution, 2-methyl was added dropwise Potassium propane-2-oleate (1M, 22.44 mL) was added for 5 minutes, anhydrous ethyl acetate (1.65 g, 18.70 mmol, 1.83 mL) was added to the mixture, and stirring was continued for 15 minutes at 0°C. The ice bath was removed, and the resulting mixture was stirred at room temperature for 2 hours. TLC showed the formation of spots, the reaction was cooled to 0°C, treated with 1.0N HCl via dropwise addition until the suspension was completely dissolved, and extracted with EtOAc (2×40 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain (Z)-3-[4-bromo-1-(tertiary butoxycarbonylamino)imidazole-2- Yl]-3-hydroxy-prop-2-enoic acid ethyl ester (2.26 g, crude product) as a yellow solid. LC-MS: m/z 376.1 [M+H] ⁺ .

Step 3

-7-羧酸乙酯(1.72g，粗產物)，為黃色固體。LC-MS：m/z 286.1[M+H]⁺。 (Z)-3-[4-Bromo-1-(tertiary butoxycarbonylamino)imidazol-2-yl]-3-hydroxy-prop-2-enoic acid ethyl ester 5d (2.26g, 6.01mmol) To the DCM (10 mL) solution, 1,1-dimethoxy-N,N-dimethyl-methylamine (2.86 g, 24.03 mmol, 3.22 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was concentrated to obtain 2-bromo-8-hydroxy-imidazo[1,2-b]

Ethyl -7-carboxylate (1.72 g, crude product) as a yellow solid. LC-MS: m/z 286.1 [M+H] ⁺ .

Step 4

-7-羧酸乙酯6d(1.72g，6.01mmol)之1,4-二

烷(20mL)攪拌溶液中，添加磷醯溴(4.31g，15.03mmol，1.53mL)。於110℃攪拌反應混合物3小時，反應完全後，冷卻反應混合物至室溫，然後將其傾入水中，以EtOAc(2×50mL)萃取。合併之有機相以鹽水(50mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮，得到粗產物，利用快速管柱層析法(石油醚/乙酸乙酯，10：1)將其純化，得到2,8-二溴咪唑并[1,2-b]嗒

-7-羧酸乙酯(1.22g，3.50mmol，58.15%產率)，為黃色固體。LC-MS：m/z 348.1[M+H]⁺。 At room temperature, in 2-bromo-8-hydroxy-imidazo[1,2-b]

-7-carboxylic acid ethyl ester 6d (1.72g, 6.01mmol) of 1,4-bis

To the stirred solution of alkane (20 mL), phosphatidyl bromide (4.31 g, 15.03 mmol, 1.53 mL) was added. The reaction mixture was stirred at 110°C for 3 hours. After the reaction was complete, the reaction mixture was cooled to room temperature, then poured into water, and extracted with EtOAc (2×50 mL). The combined organic phase was washed with brine (50 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to obtain the crude product, which was purified by flash column chromatography (petroleum ether/ethyl acetate, 10:1) , Get 2,8-dibromoimidazo[1,2-b]

Ethyl -7-carboxylate (1.22 g, 3.50 mmol, 58.15% yield) as a yellow solid. LC-MS: m/z 348.1 [M+H] ⁺ .

Step 5

-7-羧酸乙酯7d(200mg，573.10μmol)、環丙基硼酸(147.7mg，1.72mmol)之甲苯(5mL)溶液中，添加三環己膦(tricyclohexylphosphane)(160.7mg，573.10μmol)、磷酸三鉀(304.1mg，1.43mmol)與二乙醯氧鈀(25.7mg，114.62μmol)。於100℃攪拌懸浮液4小時。通過矽藻土墊過濾該混合物，濾液以水(5mL)稀釋並以乙酸乙酯(10mL)萃取。有機相以鹽水洗滌，乾燥，濃縮得到殘留物，利用管柱層析法以石油醚/乙酸乙酯(8/1)洗提將其純化，得到2-溴-8-環丙基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(118mg，380.47μmol，66.39%產率)，為白色固體。LC-MS：m/z 310.1[M+H]⁺。 At 2,8-dibromoimidazo[1,2-b]ta

-7-carboxylic acid ethyl ester 7d (200mg, 573.10 μ mol), cyclopropylboronic acid (147.7mg, 1.72mmol) in toluene (5mL) solution, add tricyclohexylphosphane (160.7mg, 573.10 μ mol) ), tripotassium phosphate (304.1 mg, 1.43 mmol) and palladium diacetyl oxide (25.7 mg, 114.62 μ mol). The suspension was stirred at 100°C for 4 hours. The mixture was filtered through a pad of Celite, and the filtrate was diluted with water (5 mL) and extracted with ethyl acetate (10 mL). The organic phase was washed with brine, dried, and concentrated to obtain a residue, which was purified by column chromatography with petroleum ether/ethyl acetate (8/1) to obtain 2-bromo-8-cyclopropyl-imidazo [1,2-b]

7-carboxylate (118mg, 380.47 μ mol, 66.39 % yield) as a white solid. LC-MS: m/z 310.1 [M+H] ⁺ .

Step 6

-7-羧酸乙酯8f(118mg，380.47μmol)之乙醇(4mL)溶液中，添加氫氧化鋰一水合物(1M，1.90mL)。於室溫攪拌混合物10小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以H₂O(10mL)稀釋該混合物並以乙酸乙酯(10mL x 2)萃取。合併之有機層以鹽水(10mL)洗 滌，以無水硫酸鈉乾燥，過濾及濃縮，得到2-溴-8-環丙基-咪唑并[1,2-b]嗒

-7-羧酸(107mg，粗產物)，為白色固體。LC-MS：m/z 287[M+H]⁺。 0℃, at 2-bromo-8-cyclopropyl-imidazo[1,2-b]

7-carboxylate 8f (118mg, 380.47 μ mol) with ethanol (4mL) was added lithium hydroxide monohydrate (1M, 1.90mL). The mixture was stirred at room temperature for 10 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was diluted with H ₂ O (10 mL) and extracted with ethyl acetate (10 mL x 2). The combined organic layer was washed with brine (10 mL), dried over anhydrous sodium sulfate, filtered and concentrated to obtain 2-bromo-8-cyclopropyl-imidazo[1,2-b]

-7-carboxylic acid (107 mg, crude product) as a white solid. LC-MS: m/z 287 [M+H] ⁺ .

Step 7

-7-羧酸9e(107mg，379.31μmol)之1,4-二

烷(7mL)溶液中，添加三乙胺(191.9mg，1.90mmol，264.34uL)與[疊氮(苯氧)磷醯基]氧苯(125.3mg，455.2μmol，98.63uL)。攪拌該混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(96.4mg，493.10μmol)，於100℃攪拌反應混合物2小時。濃縮混合物得到殘留物，利用製備型TLC(二氯甲烷：甲醇=20：1)將其純化，得到1-(2-溴-8-環丙基-咪唑并[1,2-b]嗒

-7-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(37.1mg，78.15μmol，20.60%產率)，為黃色固體。LC-MS：(ESI)m/z 474[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆)δ 9.86(s,1H),9.04(s,1H),8.71(s,1H),8.58(s,1H),8.48(s,1H),8.38(s,1H),8.16(d,J=2.0Hz,2H),2.21-2.14(m,1H),1.76-1.72(m,2H),1.17-1.14(m,2H)。 At room temperature, in 2-bromo-8-cyclopropyl-imidazo[1,2-b]

-7-carboxylic acid 9e (107mg, 379.31 μ mol) of 1,4-bis

To the alkane (7 mL) solution, triethylamine (191.9 mg, 1.90 mmol, 264.34 uL) and [azido(phenoxy) phosphatidyl] oxybenzene (125.3 mg, 455.2 μ mol, 98.63 uL) were added. The mixture was stirred for 30 minutes, then 5-chloro-6- (triazol-2-yl) pyridin-3-amine 10a (96.4mg, 493.10 μ mol) , stirring the reaction mixture for 2 hours at 100 deg.] C. The mixture was concentrated to obtain a residue, which was purified by preparative TLC (dichloromethane: methanol = 20:1) to obtain 1-(2-bromo-8-cyclopropyl-imidazo[1,2-b]

7-yl) -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (37.1mg, 78.15 μ mol, 20.60 % yield) as a yellow solid. LC-MS: (ESI) m/z 474 [M+H] ⁺ . ¹ H NMR(400MHz, DMSO- d ₆ ) δ 9.86(s,1H), 9.04(s,1H), 8.71(s,1H), 8.58(s,1H), 8.48(s,1H), 8.38(s , 1H), 8.16 (d, J = 2.0 Hz, 2H), 2.21-2.14 (m, 1H), 1.76-1.72 (m, 2H), 1.17-1.14 (m, 2H).

-7-基)脲之合成 Example 8. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-cyclopropylimidazo[1,2 -b] Da

-7-yl) urea synthesis

step 1

At 0°C, in 1-((tertiary butoxycarbonyl)amino)-1H-imidazole-2-carboxylic acid ethyl ester 3 (1.5g, 5.882mmol) in THF (4.5mL) solution, add dropwise cold (Pre-cooled at 0°C) potassium tertiary butoxide (1M, 17.6 mL) for 5 minutes. Anhydrous ethyl acetate (1.29 g, 14.7 mmol, 1.43 mL) pre-cooled at 0°C was added dropwise, and stirring was continued at 0°C for 15 minutes. The ice bath was removed, and the resulting mixture was stirred at room temperature for 2 hours. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was cooled to 0°C, and acidified with 1.0 N HCl via dropwise addition until the suspension was completely dissolved. Extract with ethyl acetate (2×40 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The resulting residue was purified by flash column chromatography (eluted with 0% to 20% ethyl acetate in petroleum) to obtain (Z)-3-(1-((tertiary butoxycarbonyl)amino) -1H-imidazol-2-yl)-3-hydroxyethyl acrylate (380 mg, 21.8% yield), as a pale yellow solid. LC-MS: m/z 298 [M+H] ⁺ .

Step 2

-7-羧酸乙酯(122mg，45.9%產率)，為黃色油-固體。LC-MS：m/z 208[M+H]⁺。 (Z)-3-(1-((tertiary butoxycarbonyl)amino)-1H-imidazol-2-yl)-3-hydroxyethyl acrylate 5a (380mg, 1.279mmol) in DCM (10mL) , 1,1-Dimethoxy-N,N-dimethyl-methylamine (762 mg, 6.397 mmol) was added. The reaction mixture was stirred at room temperature for 13 hours. The starting material was consumed and the required quality was detected by LC-MS. The mixture was concentrated, and the residue was purified by flash chromatography (eluted with methanol/dichloromethane=10/1) to obtain 8-hydroxyimidazo[1,2-b]

Ethyl -7-carboxylate (122 mg, 45.9% yield), yellow oil-solid. LC-MS: m/z 208 [M+H] ⁺ .

Step 3

-7-羧酸乙酯6a(122mg，0.5894mmol)之1,4-二

烷(8mL)溶液中，添加磷醯溴(506.9mg，1.768mmol)。於110℃攪拌反應混合物2小時。耗盡起始原料，並以LC-MS檢測到所需質量。冷卻反應混合物至室溫，傾入水中，以乙酸乙酯(2×15mL)萃取。合併之有機相以鹽水(10mL)洗滌，以無水Na₂SO₄乾燥，過濾及濃縮。粗產物利用快速管柱層析法(以於石油中1%至25%之乙酸乙酯洗提)純化，得到8-溴咪唑并[1,2-b]嗒

-7-羧酸乙酯(110mg，69.4%產率)，為白色固體。LC-MS：m/z 270[M+H]⁺。 At room temperature, in 8-hydroxyimidazo[1,2-b]

-7-carboxylic acid ethyl ester 6a (122mg, 0.5894mmol) of 1,4-bis

To the alkane (8 mL) solution, phosphatidyl bromide (506.9 mg, 1.768 mmol) was added. The reaction mixture was stirred at 110°C for 2 hours. The starting material was consumed, and the required quality was detected by LC-MS. The reaction mixture was cooled to room temperature, poured into water, and extracted with ethyl acetate (2×15 mL). The combined organic phase was washed with brine (10 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. The crude product was purified by flash column chromatography (eluted with 1% to 25% ethyl acetate in petroleum) to obtain 8-bromoimidazo[1,2-b]

Ethyl -7-carboxylate (110 mg, 69.4% yield), a white solid. LC-MS: m/z 270 [M+H] ⁺ .

Step 4

-7-羧酸乙酯7a(110.0mg，0.4089mmol)之甲苯(5mL)溶液中，添加環丙基硼酸(70.3mg，0.8178mmol)、三環己膦(114.5mg，0.4089mmol)與磷酸鉀(216.7mg，1.022mmol)。將懸浮液脫氣並與N₂交換兩次。添加乙酸鈀(II)(18.3mg，0.08178mmol)至該混合物中。加熱反應混合物至100℃ 12小時。耗盡起始原料並以LC-MS檢測到所需質量。減壓濃縮混合物，所得殘留物以快速管柱層析法(以於石油中1%至15%之乙酸乙酯洗提)純化，得到8-環丙 基咪唑并[1,2-b]嗒

-7-羧酸乙酯(34.0mg，36%產率)，為黃色固體。LC-MS：m/z 232[M+H]⁺。 To 8-bromoimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 7a (110.0mg, 0.4089mmol) in toluene (5mL), add cyclopropylboronic acid (70.3mg, 0.8178mmol), tricyclohexylphosphine (114.5mg, 0.4089mmol) and potassium phosphate (216.7 mg, 1.022 mmol). The suspension was degassed and _{exchanged with N 2} twice. Palladium(II) acetate (18.3 mg, 0.08178 mmol) was added to the mixture. The reaction mixture was heated to 100°C for 12 hours. The starting material was consumed and the required quality was detected by LC-MS. The mixture was concentrated under reduced pressure, and the resulting residue was purified by flash column chromatography (eluted with 1% to 15% ethyl acetate in petroleum) to obtain 8-cyclopropylimidazo[1,2-b]

Ethyl -7-carboxylate (34.0 mg, 36% yield), as a yellow solid. LC-MS: m/z 232 [M+H] ⁺ .

Step 5

-7-羧酸乙酯8g(67mg，0.29mmol)之THF(4mL)溶液中，添加氫氧化鋰一水合物(30.4mg，0.7251mmol)之水(0.7mL)溶液。於30℃攪拌混合物4小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮去除溶劑，殘留物以HCl(1M)酸化至pH=3~4。以乙酸乙酯萃取該混合物並乾燥有機相，濃縮，得到8-環丙基咪唑并[1,2-b]嗒

-7-羧酸(55mg，93.4%產率)，為白色固體。LC-MS：m/z 204[M+H]⁺。 On 8-cyclopropylimidazo[1,2-b]ta

To a solution of 8 g (67 mg, 0.29 mmol) of ethyl -7-carboxylate in THF (4 mL), a solution of lithium hydroxide monohydrate (30.4 mg, 0.7251 mmol) in water (0.7 mL) was added. The mixture was stirred at 30°C for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. Concentrate to remove the solvent, and the residue is acidified with HCl (1M) to pH=3~4. The mixture was extracted with ethyl acetate and the organic phase was dried and concentrated to obtain 8-cyclopropylimidazo[1,2-b]

-7-carboxylic acid (55 mg, 93.4% yield), as a white solid. LC-MS: m/z 204 [M+H] ⁺ .

Step 6

-7-羧酸9f(55mg，0.2709mmol)之1,4-二

烷(4mL)溶液中，添加疊氮膦酸二苯酯(89.5mg，0.3251mmol)與三乙胺(137.1mg，1.354mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(79.2mg，0.4064mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。該有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(8-環丙基咪唑并[1,2-b]嗒

-7-基)脲(9.1mg)，為白色固體。LC-MS：m/z 396[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.73(s,1 H),8.93(s,1 H),8.58-8.57(m,2 H),8.47(d,J=2.4Hz,1 H),8.15(s,2 H),8.13(d,J=1.2Hz,1 H),7.61(d,J=1.2Hz,1 H),2.22-2.18(m,1 H),1.89-1.86(m,2 H),1.15-1.10(m,2 H)。 On 8-cyclopropylimidazo[1,2-b]ta

-7-carboxylic acid 9f (55mg, 0.2709mmol) of 1,4-bis

To the alkane (4 mL) solution, diphenyl azide phosphonate (89.5 mg, 0.3251 mmol) and triethylamine (137.1 mg, 1.354 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (79.2 mg, 0.4064 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The residue obtained was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(5-chloro-6- (2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-cyclopropylimidazo[1,2-b]ta

-7-yl)urea (9.1 mg) as a white solid. LC-MS: m/z 396 [M+H] ⁺ . ¹ H NMR (DMSO- d6 ,400MHz): δ 9.73 (s, 1 H), 8.93 (s, 1 H), 8.58-8.57 (m, 2 H), 8.47 (d, J = 2.4 Hz, 1 H) , 8.15 (s, 2 H), 8.13 (d, J = 1.2 Hz, 1 H), 7.61 (d, J = 1.2 Hz, 1 H), 2.22-2.18 (m, 1 H), 1.89-1.86 (m , 2 H), 1.15-1.10 (m, 2 H).

-7-基)脲之合成 Example 9. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-propylimidazo [1,2- b]

-7-yl) urea synthesis

step 1

-7-羧酸9a(60mg，0.2532mmol)之甲醇(3mL)溶液中，添加Pd/C(30mg)。H₂下，於室溫攪拌反應混合物3小時。耗盡起始原料並以LC-MS檢測到所需質量。將其過濾並減壓濃縮濾液。粗產物利用快速管柱層析法(以於石油中0%至80%之乙酸乙酯洗提)純化，得到8-丙基咪唑并[1,2-b]嗒

-7-羧酸(50mg,96.3%產率)，為白色固體。LC-MS：m/z 206[M+H]⁺。 On 2-chloro-8-cyclopropylimidazo[1,2-b]

To a solution of -7-carboxylic acid 9a (60 mg, 0.2532 mmol) in methanol (3 mL), Pd/C (30 mg) was added. Under H _2, the reaction mixture was stirred at room temperature for 3 hours. The starting material was consumed and the required quality was detected by LC-MS. It was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (eluted with 0% to 80% ethyl acetate in petroleum) to obtain 8-propylimidazo[1,2-b]

-7-carboxylic acid (50 mg, 96.3% yield), as a white solid. LC-MS: m/z 206 [M+H] ⁺ .

Step 2

-7-羧酸9g(50mg，0.2439mmol)之1,4-二

烷(4mL)溶液中，添加疊氮膦酸二苯酯(87.2mg，0.3171mmol)與三乙胺(123.4mg，1.219mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(71.3mg，0.366mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。該有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(8-丙基咪唑并[1,2-b]嗒

-7-基)脲(15.7mg)，為白色固體。LC-MS：m/z 398[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.75(s,1 H),8.77(s,1 H),8.76(s,1 H),8.57(d,J=2.4Hz,1 H),8.48(d,J=2.4Hz,1 H),8.18(d,J=1.2Hz,1 H),8.15(s,2 H),7.67(d,J=1.6Hz,1 H),2.98-2.94(m,2 H),1.73-1.69(m,2 H),0.99-0.96(m,3 H)。 On 8-propylimidazo[1,2-b]

-7-carboxylic acid 9g (50mg, 0.2439mmol) of 1,4-bis

To the alkane (4 mL) solution, add diphenyl azide phosphonate (87.2 mg, 0.3171 mmol) and triethylamine (123.4 mg, 1.219 mmol). The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (71.3 mg, 0.366 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The residue obtained was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(5-chloro-6- (2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-propylimidazo[1,2-b]ta

-7-yl)urea (15.7 mg) as a white solid. LC-MS: m/z 398 [M+H] ⁺ . ¹ H NMR (DMSO- d6 ,400MHz): δ 9.75(s,1 H), 8.77(s,1 H), 8.76(s,1 H), 8.57(d, J =2.4Hz,1 H), 8.48 (d, J =2.4Hz,1 H), 8.18(d, J =1.2Hz,1 H), 8.15(s, 2 H), 7.67(d, J =1.6Hz,1 H), 2.98-2.94( m, 2 H), 1.73-1.69 (m, 2 H), 0.99-0.96 (m, 3 H).

-7-基)脲之合成 Example 10. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloroimidazo[1,2-b ]despair

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(50mg，0.165mmol)之乙醇(8mL)溶液中，添加Pd/C(5mg)。H₂下，於室溫攪拌反應混合物3小時。耗盡起始原料並以LC-MS檢測到所需質量。將其過濾並減壓濃縮濾液。粗產物利用快速管柱層析法(以於石油中0%至30%之乙酸乙酯洗提)純化，得到2-氯咪唑并[1,2-b]嗒

-7-羧酸乙酯(25mg，67.4%產率)，為白色固體。LC-MS：m/z 226[M+H]⁺。 On 8-bromo-2-chloroimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 7c (50 mg, 0.165 mmol) in ethanol (8 mL), Pd/C (5 mg) was added. Under H _2, the reaction mixture was stirred at room temperature for 3 hours. The starting material was consumed and the required quality was detected by LC-MS. It was filtered and the filtrate was concentrated under reduced pressure. The crude product was purified by flash column chromatography (eluted with 0% to 30% ethyl acetate in petroleum) to obtain 2-chloroimidazo[1,2-b]

Ethyl -7-carboxylate (25 mg, 67.4% yield), a white solid. LC-MS: m/z 226 [M+H] ⁺ .

Step 2

-7-羧酸乙酯15c(42mg，0.1867mmol)之THF(2mL)溶液中，添加氫氧化鋰一水合物(15.7mg，0.373mmol)之水(0.3mL)溶液。於30℃攪拌混合物4小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮所得混合物以移除溶劑，殘留物以HCl(1M)酸化至pH=3~4。以乙酸乙酯萃取該混合物並乾燥有機相，濃縮，得到2-氯咪唑并[1,2-b]嗒

-7-羧酸(35mg，95.1%)，為黃色固體。LC-MS：m/z 198[M+H]⁺。 At 2-chloroimidazo[1,2-b]

To a THF (2 mL) solution of ethyl-7-carboxylate 15c (42 mg, 0.1867 mmol), a water (0.3 mL) solution of lithium hydroxide monohydrate (15.7 mg, 0.373 mmol) was added. The mixture was stirred at 30°C for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. The resulting mixture was concentrated to remove the solvent, and the residue was acidified with HCl (1M) to pH=3~4. The mixture was extracted with ethyl acetate and the organic phase was dried and concentrated to give 2-chloroimidazo[1,2-b]

-7-carboxylic acid (35 mg, 95.1%), as a yellow solid. LC-MS: m/z 198[M+H] ⁺ .

Step 3

-7-羧酸16c(35mg，0.1777mmol)之1,4-二

烷(4mL)溶液中，添加疊氮膦酸二苯酯(58.7mg，0.213mmol)與三乙胺(89.9mg，0.8885mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(52mg，0.267mmol)，加熱混合物至100℃ 2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。該有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至100%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯咪唑并[1,2-b]嗒

-7-基)脲(10.2mg)，為白色固體。LC-MS：m/z 390[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.87(s,1 H),9.75(s,1H),8.63-8.61(m,2 H),8.46(d,J=2.4Hz,1 H),8.34(s,1 H),8.16-8.13(m,3 H)。 At 2-chloroimidazo[1,2-b]

-7-carboxylic acid 16c (35mg, 0.1777mmol) of 1,4-bis

To the alkane (4 mL) solution, diphenyl azide phosphonate (58.7 mg, 0.213 mmol) and triethylamine (89.9 mg, 0.8885 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (52 mg, 0.267 mmol) was added, and the mixture was heated to 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The resulting residue was purified by flash column chromatography (eluted with 10% to 100% ethyl acetate in petroleum) to obtain 1-(5-chloro-6- (2H-1,2,3-Triazol-2-yl)pyridin-3-yl)-3-(2-chloroimidazo[1,2-b]ta

-7-yl)urea (10.2 mg) as a white solid. LC-MS: m/z 390 [M+H] ⁺ . ¹ H NMR(DMSO- d6 ,400MHz): δ 9.87(s,1 H),9.75(s,1H),8.63-8.61(m,2 H),8.46(d, J =2.4Hz,1 H), 8.34 (s, 1 H), 8.16 to 8.13 (m, 3 H).

-7-基)脲之合成 Example 11. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2,8-dichloroimidazo[1, 2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯6c(200mg，0.8299mmol)之THF(4mL)溶液中，添加氫氧化鋰一水合物(69.7mg，1.66mmol)之水(1.6mL)溶液。於70℃攪拌混合物14小時。耗盡起始原料並以LC-MS檢測到所需質量。 濃縮去除溶劑，殘留物以HCl(1M)酸化至pH=3~4。以乙酸乙酯萃取該混合物並乾燥有機相，濃縮，得到2-氯-8-羥基咪唑并[1,2-b]嗒

-7-羧酸(160mg，90.5%)，為白色固體。LC-MS：m/z 214[M+H]⁺。 On 2-chloro-8-hydroxyimidazo[1,2-b]ta

To a solution of ethyl -7-carboxylate 6c (200 mg, 0.8299 mmol) in THF (4 mL), a solution of lithium hydroxide monohydrate (69.7 mg, 1.66 mmol) in water (1.6 mL) was added. The mixture was stirred at 70°C for 14 hours. The starting material was consumed and the required quality was detected by LC-MS. Concentrate to remove the solvent, and the residue is acidified with HCl (1M) to pH=3~4. The mixture was extracted with ethyl acetate and the organic phase was dried and concentrated to give 2-chloro-8-hydroxyimidazo[1,2-b]

-7-carboxylic acid (160 mg, 90.5%), as a white solid. LC-MS: m/z 214 [M+H] ⁺ .

Step 2

-7-羧酸9h(187.0mg，929.1μmol)之POCl₃(6mL)溶液12小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮反應液，殘留物以水(10mL)猝滅並以DCM(25mL x 2)萃取。有機層以鹽水(20mL)洗滌，以無水Na₂SO₄乾燥並減壓濃縮，殘留物利用快速層析法(以甲醇/二氯甲烷=10/1洗提)純化，得到2,8-二氯咪唑并[1,2-b]嗒

-7-羧酸(150mg，粗產物)之粗產物，為白色固體。LC-MS：m/z 232[M+H]⁺。 Heat 2-chloro-8-hydroxyimidazo[1,2-b]ta at 120℃

-7-carboxylic acid 9h (187.0 mg, 929.1 μmol) in POCl ₃ (6 mL) solution for 12 hours. The starting material was consumed and the required quality was detected by LC-MS. The reaction solution was concentrated, and the residue was quenched with water (10 mL) and extracted with DCM (25 mL x 2). The organic layer was washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ and concentrated under reduced pressure. The residue was purified by flash chromatography (eluted with methanol/dichloromethane = 10/1) to obtain 2,8-di Chlorimidazo[1,2-b]ta

The crude product of -7-carboxylic acid (150 mg, crude product) was a white solid. LC-MS: m/z 232 [M+H] ⁺ .

Step 3

-7-羧酸9i(50mg，0.2164mmol)之1,4-二

烷(5mL)溶液中，添加疊氮膦酸二苯酯(71.5mg，0.2597mmol)與三乙胺(109.5mg，1.082mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(63.3mg，0.3246mmol)，混合物於100℃加熱2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至100%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2,8-二氯咪唑并[1,2-b]嗒

-7-基)脲(3.5mg)，為白色固體。LC-MS：m/z 424[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 10.11(s,1 H),9.25(s,2 H),8.57(d,J=2.4Hz,1 H),8.52(s,1 H),8.48(d,J=2.4Hz,1 H),8.16(s,2 H)。 At 2,8-Dichloroimidazo[1,2-b]ta

-7-carboxylic acid 9i (50mg, 0.2164mmol) of 1,4-bis

To the alkane (5 mL) solution, add diphenyl azide phosphonate (71.5 mg, 0.2597 mmol) and triethylamine (109.5 mg, 1.082 mmol). The resulting mixture was stirred at room temperature for 30 minutes, then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (63.3 mg, 0.3246 mmol) was added, and the mixture was heated at 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The resulting residue was purified by flash column chromatography (eluted with 10% to 100% ethyl acetate in petroleum) to obtain 1-(5-chloro-6-( 2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2,8-dichloroimidazo[1,2-b]ta

-7-yl)urea (3.5 mg) as a white solid. LC-MS: m/z 424 [M+H] ⁺ . ¹ H NMR(DMSO- d6 ,400MHz): δ 10.11(s,1 H), 9.25(s, 2 H), 8.57(d, J =2.4Hz,1 H), 8.52(s,1 H), 8.48 (d, J = 2.4 Hz, 1 H), 8.16 (s, 2 H).

-7-基)脲之合成 Example 12. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-methylimidazo[ 1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(200mg，656.76μmol)、甲基硼酸(147.4mg，2.46mmol)與三環己膦(230.2mg，820.94μmol)之1,4-二

烷(5mL)溶液中，添加磷酸鉀(522.8mg，2.46mmol)與乙酸鈀(II)(92.2mg，410.47μmol)。N₂下，於100℃攪拌懸浮液5小時。冷卻至室溫後，過濾反應混合物，濃縮濾液，得到殘留物，利用快速管柱層析法(以PE/EA=10/1洗提)將其純化，得到2-氯-8-甲基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(68mg，283.74μmol，43.20%產率)，為白色固體。LC-MS：m/z 240[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

-7-carboxylic acid ethyl ester 7c (200mg, 656.76 μ mol), methylboronic acid (147.4mg, 2.46mmol) and tricyclohexylphosphine (230.2mg, 820.94 μ mol) of 1,4-bis

Dioxane (5mL) was added potassium phosphate (522.8mg, 2.46mmol) and palladium acetate (II) (92.2mg, 410.47 μ mol). The suspension was stirred at 100°C for 5 hours under _{N 2.} After cooling to room temperature, the reaction mixture was filtered, and the filtrate was concentrated to obtain the residue, which was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-methyl- Imidazo[1,2-b]

7-carboxylate (68mg, 283.74 μ mol, 43.20 % yield) as a white solid. LC-MS: m/z 240 [M+H] ⁺ .

Step 2

-7-羧酸乙酯8h(85mg，354.67μmol)之THF(3mL)溶液中，添加氫氧化鋰一水合物(29.8mg，709.34μmol)之水(0.7mL)溶液。於室溫攪拌混合物5小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取該混合物，乾燥有機相，濃縮，得到2-氯-8-甲基-咪唑并[1,2-b]嗒

-7-羧酸(74mg，99%產率)，為白色固體。LC-MS：m/z 212[M+H]⁺。 On 2-chloro-8-methyl-imidazo[1,2-b]

To the 8h (85 mg, 354.67 μ mol) of ethyl -7-carboxylate in THF (3 mL) was added lithium hydroxide monohydrate (29.8 mg, 709.34 μ mol) in water (0.7 mL). The mixture was stirred at room temperature for 5 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc, the organic phase was dried and concentrated to give 2-chloro-8-methyl-imidazo[1,2-b]

-7-carboxylic acid (74 mg, 99% yield), as a white solid. LC-MS: m/z 212 [M+H] ⁺ .

Step 3

烷(6mL)溶液中，添加疊氮膦酸二苯酯(108.8mg，419.65μmol)與三乙胺(176.9mg，1.75mmol)。所得溶液於室溫攪拌30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(102.6mg，524.56μmol)，於100℃攪拌該混合物2小時。去除溶劑，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)與製備型HPLC純化，得到1-(2-氯-7-甲基-吡唑并[1,5-a]嘧啶-6-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(33mg，23%產率)。LC-MS：m/z 403.8。¹H NMR(DMSO,400MHz)：δ 9.79-9.12(m,2H),8.89(s,1H),8.58(d,J=1.6Hz,1H),8.48(d,J=2.0Hz,1H),8.37(s,1H),8.16(s,2H),2.47(s,3H)。 In 2-chloro-7-methyl-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid 9j (74mg, 349.71 μ mol) of 1,4-bis

Dioxane (6mL) was added azide diphenyl phosphonate (108.8mg, 419.65 μ mol) and triethylamine (176.9mg, 1.75mmol). The resulting solution was stirred at room temperature for 30 minutes and then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine 10a (102.6mg, 524.56 μ mol) , and the mixture was stirred at 100 ℃ 2 hours. The solvent was removed, and the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) and preparative HPLC to obtain 1-(2-chloro-7-methyl-pyrazolo[1,5 -a]pyrimidin-6-yl)-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (33 mg, 23% yield). LC-MS: m/z 403.8. ¹ H NMR(DMSO,400MHz): δ 9.79-9.12(m,2H), 8.89(s,1H), 8.58(d,J=1.6Hz,1H), 8.48(d,J=2.0Hz,1H), 8.37 (s, 1H), 8.16 (s, 2H), 2.47 (s, 3H).

Synthesis of compound of formula II

-7-基)脲之合成 Example 13. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-methoxyimidazo [1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯6c(100mg，0.41mmol)與碳酸鉀(171mg，1.24mmol)之DMF(5mL)攪拌溶液中，添加碘甲烷(176.2mg，1.24mmol，77.29uL)。於室溫攪拌反應混合物2小時，反應完全後，真空 濃縮該混合物。粗產物利用快速管柱層析法(SiO₂,DCM/MeOH=10：1)純化，得到2-氯-8-甲氧基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(90mg，352.03μmol，85%產率)，為黃色固體。LC-MS：m/z 256.1[M+H]⁺。 At room temperature, in 2-chloro-8-hydroxy-imidazo[1,2-b]

To a stirred solution of ethyl -7-carboxylate 6c (100 mg, 0.41 mmol) and potassium carbonate (171 mg, 1.24 mmol) in DMF (5 mL), methyl iodide (176.2 mg, 1.24 mmol, 77.29 uL) was added. The reaction mixture was stirred at room temperature for 2 hours. After the reaction was complete, the mixture was concentrated in vacuo. The crude product was purified by flash column chromatography (SiO ₂ ,DCM/MeOH=10:1) to obtain 2-chloro-8-methoxy-imidazo[1,2-b]

7-carboxylate (90mg, 352.03 μ mol, 85 % yield) as a yellow solid. LC-MS: m/z 256.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯11a(90mg，352.03μmol)之EtOH(2mL)與H₂O(2mL)攪拌溶液中，添加LiOH H₂O(73.9mg，1.76mmol)。於室溫攪拌反應混合物3小時，反應完全後，以EtOAc(5mL)稀釋該混合物並以1N HCl調至pH=2。分離各層，水層以EtOAc(2×5mL)萃取。合併之有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物2-氯-8-甲氧基-咪唑并[1,2-b]嗒

-7-羧酸(73mg，320.73μmol，91%產率)為黃色固體，直接用於下一步驟。LC-MS：m/z 226.0[M-H]⁺。 On 2-chloro-8-methoxy-imidazo[1,2-b]

7-carboxylate 11a (90mg, 352.03 μ mol) of EtOH (2mL) and H ₂ O (2mL) was stirred solution was added _{LiOH H 2 O (73.9mg, 1.76mmol} ). The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was diluted with EtOAc (5 mL) and adjusted to pH=2 with 1N HCl. The layers were separated, and the aqueous layer was extracted with EtOAc (2×5 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. Crude product 2-chloro-8-methoxy-imidazo[1,2-b]

-7-carboxylic acid (73 mg, 320.73 μ mol, 91% yield) was a yellow solid, which was used directly in the next step. LC-MS: m/z 226.0 [MH] ⁺ .

Step 3

-7-羧酸12a(65mg，285.54μmol)之1,4-二

烷(4mL)攪拌溶液中，添加二苯基磷醯基疊氮化物(102.2mg，371.20μmol，80.44uL)、Et₃N(1.43mmol)。於室溫攪拌反應混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(111.7mg，571.08μmol)，於100℃攪拌混合物2小時，反應完全後，以EtOAc(4mL)稀釋該混合物，有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物利用快速管柱層析法(SiO₂,DCM/MeOH=20：1)純化，得到1-(2-氯-8-甲氧基-咪唑并[1,2-b]嗒

-7-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(10.3mg，24.51μmol，8%產率)，為白色固體。LC-MS：m/z 408.0[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆ )：10.27(s,1H),9.15(s,1H),8.73(s,1H),8.49-8.46(m,3H),8.15(s,2H),4.25(s,3H)。 On 2-chloro-8-methoxy-imidazo[1,2-b]

-7-carboxylic acid 12a (65mg, 285.54 μ mol) of 1,4-bis

Dioxane (4mL) stirred solution was added diphenylphosphoryl acyl azide (102.2mg, 371.20 μ mol, 80.44uL ), Et 3 N (1.43mmol). The reaction mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (111.7 mg, 571.08 μ mol) was added, and the mixture was stirred at 100°C for 2 hours. The reaction was complete Afterwards, the mixture was diluted with EtOAc (4 mL), and the organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (SiO ₂ ,DCM/MeOH=20:1) to obtain 1-(2-chloro-8-methoxy-imidazo[1,2-b]

7-yl) -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (10.3mg, 24.51 μ mol, 8 % yield) as a white solid. LC-MS: m/z 408.0 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO- d ₆ ): 10.27 (s, 1H), 9.15 (s, 1H), 8.73 (s, 1H), 8.49-8.46 (m, 3H), 8.15 (s, 2H), 4.25 (s,3H).

-7-基)脲之合成 Example 14. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-ethoxyimidazo [1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯6c(200mg，827.71μmol)之DMF(5mL)攪拌溶液中，添加碘乙烷(387.3mg，2.48mmol，199.63uL)。微波輻射下，於80℃攪拌反應混合物2小時，反應完全後，真空濃縮混合物。粗產物利用快速管柱層析法(SiO₂,DCM/MeOH=10：1)純化，得到2-氯-8-乙氧基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(103mg，381.93μmol，46%產率)，為黃色固體。LC-MS：m/z 270.1[M+H]⁺。 At room temperature, in 2-chloro-8-hydroxy-imidazo[1,2-b]

7-carboxylic acid ethyl ester 6c (200mg, 827.71 μ mol) of DMF (5mL) was stirred solution was added ethyl iodide (387.3mg, 2.48mmol, 199.63uL). Under microwave irradiation, the reaction mixture was stirred at 80°C for 2 hours. After the reaction was completed, the mixture was concentrated in vacuo. The crude product was purified by flash column chromatography (SiO ₂ , DCM/MeOH=10:1) to obtain 2-chloro-8-ethoxy-imidazo[1,2-b]

7-carboxylate (103mg, 381.93 μ mol, 46 % yield) as a yellow solid. LC-MS: m/z 270.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯11b(103mg，381.93μmol)之EtOH(2mL)與H₂O(2mL)攪拌溶液中，添加LiOH H₂O(73.9mg，1.76mmol)。於室溫攪拌反應混合物3小時，反應完全後，以EtOAc(5mL)稀釋該混合物並以1N HCl調至pH=2。分離各層，水層以EtOAc(2×5mL)萃取。合併之有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物2-氯-8-乙氧 基-咪唑并[1,2-b]嗒

-7-羧酸(70mg，289.70μmol，76%產率)為黃色固體，直接用於下一步驟。LC-MS：m/z 240.0[M-H]⁺。 On 2-chloro-8-ethoxy-imidazo[1,2-b]ta

7-carboxylate 11b (103mg, 381.93 μ mol) of EtOH (2mL) and H ₂ O (2mL) was stirred solution was added _{LiOH H 2 O (73.9mg, 1.76mmol} ). The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was diluted with EtOAc (5 mL) and adjusted to pH=2 with 1N HCl. The layers were separated, and the aqueous layer was extracted with EtOAc (2×5 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. Crude product 2-chloro-8-ethoxy-imidazo[1,2-b]

7-carboxylic acid (70mg, 289.70 μ mol, 76 % yield) as a yellow solid which was used directly in the next step. LC-MS: m/z 240.0 [MH] ⁺ .

Step 3

-7-羧酸12b(70mg，289.70μmol)之1,4-二

烷(4mL)攪拌溶液中，添加二苯基磷醯基疊氮化物(159.5mg，579.40μmol，125.55uL)、Et₃N(146.3mg，1.45mmol)。於室溫攪拌反應混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(113.3mg，579.40μmol)，於100℃攪拌混合物2小時，反應完全後，以EtOAc(4mL)稀釋該混合物，有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物利用快速管柱層析法(SiO₂,DCM/MeOH=20：1)純化，得到1-(2-氯-8-乙氧基-咪唑并[1,2-b]嗒

-7-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(8.6mg，19.80μmol，6.84%產率)，為白色固體。LC-MS：m/z 434.0[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆ )：10.27-10.26(m,1H),9.14(s,1H),8.76(s,1H),8.50-8.47(m,3H),8.15(s,2H),4.79(q,J=6.8Hz,2H),1.40(t,J=6.8Hz,3H)。 On 2-chloro-8-ethoxy-imidazo[1,2-b]ta

-7-carboxylic acid 12b (70mg, 289.70 μ mol) of 1,4-bis

Dioxane (4mL) stirred solution was added diphenylphosphoryl acyl azide (159.5mg, 579.40 μ mol, 125.55uL ), Et 3 N (146.3mg, 1.45mmol). The reaction mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (113.3 mg, 579.40 μ mol) was added, and the mixture was stirred at 100°C for 2 hours. The reaction was complete Afterwards, the mixture was diluted with EtOAc (4 mL), and the organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (SiO ₂ ,DCM/MeOH=20:1) to obtain 1-(2-chloro-8-ethoxy-imidazo[1,2-b]

7-yl) -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (8.6mg, 19.80 μ mol, 6.84 % yield) as a white solid. LC-MS: m/z 434.0 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO- d ₆ ): 10.27-10.26 (m, 1H), 9.14 (s, 1H), 8.76 (s, 1H), 8.50-8.47 (m, 3H), 8.15 (s, 2H) ,4.79(q, J =6.8Hz,2H),1.40(t, J =6.8Hz,3H).

-7-基)脲之合成 Example 15. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-isopropoxyimidazole And [1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯6c(350mg，1.45mmol)與碳酸銫(1.42g，4.35mmol)之DMF(5mL)攪拌溶液中，添加2-溴丙烷(1.78g，14.48mmol，248.30uL)利用快速管柱層析法(SiO₂,DCM/MeOH=10：1)純化。微波輻射下，所得混合物於120℃攪拌2小時，反應完全後，真空濃縮混合物。粗產物利用快速管柱層析法(SiO₂,DCM/MeOH=10：1)純化，得到2-氯-8-異丙氧基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(82mg，289.03μmol，20%產率)，為黃色油。LC-MS：m/z 84.1[M+H]⁺。 At room temperature, in 2-chloro-8-hydroxy-imidazo[1,2-b]

To a stirred solution of ethyl -7-carboxylate 6c (350mg, 1.45mmol) and cesium carbonate (1.42g, 4.35mmol) in DMF (5mL), add 2-bromopropane (1.78g, 14.48mmol, 248.30uL) using rapid Purification by column chromatography (SiO ₂ , DCM/MeOH=10:1). Under microwave irradiation, the resulting mixture was stirred at 120°C for 2 hours. After the reaction was completed, the mixture was concentrated in vacuo. The crude product was purified by flash column chromatography (SiO ₂ , DCM/MeOH=10:1) to obtain 2-chloro-8-isopropoxy-imidazo[1,2-b]

7-carboxylate (82mg, 289.03 μ mol, 20 % yield) as a yellow oil. LC-MS: m/z 84.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯11c(82mg，289.03μmol)之EtOH(2mL)與H₂O(2mL)攪拌溶液中，添加LiOH H₂O(60.70mg，1.45mmol)。於室溫攪拌反應混合物3小時，反應完全後，以EtOAc(5mL)稀釋該混合物並以1N HCl調至pH=2。分離各層，水層以EtOAc(2×5mL)萃取。合併之有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物2-氯-8-異 丙氧基-咪唑并[1,2-b]嗒

-7-羧酸(67mg，262.07μmol，91%產率)為黃色固體，直接用於下一步驟。LC-MS：m/z 254.0[M-H]⁺。 On 2-chloro-8-isopropoxy-imidazo[1,2-b]

7-carboxylate 11c (82mg, 289.03 μ mol) of EtOH (2mL) was stirred with H ₂ O (2mL) was added _{LiOH H 2 O (60.70mg, 1.45mmol} ). The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was diluted with EtOAc (5 mL) and adjusted to pH=2 with 1N HCl. The layers were separated, and the aqueous layer was extracted with EtOAc (2×5 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. Crude product 2-chloro-8-isopropoxy-imidazo[1,2-b]

7-carboxylic acid (67mg, 262.07 μ mol, 91 % yield) as a yellow solid which was used directly in the next step. LC-MS: m/z 254.0 [MH] ⁺ .

Step 3

-7-羧酸12c(21.2mg，82.77μmol)之1,4-二

烷(4mL)攪拌溶液中，添加二苯基磷醯基疊氮化物(45.6mg，165.54μmol，35.87uL)、Et₃N(41.80mg，413.85μmol)。於室溫攪拌反應混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(32.4mg，165.54μmol)，於100℃攪拌混合物2小時，反應完全後，以EtOAc(4mL)稀釋該混合物，有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物利用快速管柱層析法(矽膠，DCM/MeOH=20：1)純化，得到1-(2-氯-8-異丙氧基-咪唑并[1,2-b]嗒

-7-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(11.3mg，25.21μmol，30%產率)，為白色固體。LC-MS：m/z 448.0[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆ )：10.29(s,1H),9.15(s,1H),8.78(s,1H),8.50-8.47(m,3H),8.15(s,2H),3.35-3.29(m,1H)1.68(d,J=6.4Hz,2H)。 On 2-chloro-8-isopropoxy-imidazo[1,2-b]

-7-carboxylic acid 12c (21.2mg, 82.77 μ mol) of 1,4-bis

To the alkane (4 mL) stirring solution, diphenylphosphoryl azide (45.6 mg, 165.54 μ mol, 35.87 uL) and Et ₃ N (41.80 mg, 413.85 μ mol) were added. The reaction mixture was stirred at room temperature for 30 minutes, then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (32.4 mg, 165.54 μ mol) was added, and the mixture was stirred at 100°C for 2 hours. The reaction was complete Afterwards, the mixture was diluted with EtOAc (4 mL), and the organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (silica gel, DCM/MeOH=20:1) to obtain 1-(2-chloro-8-isopropoxy-imidazo[1,2-b]

7-yl) -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (11.3mg, 25.21 μ mol, 30 % yield) as a white solid. LC-MS: m/z 448.0 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO- d ₆ ): 10.29 (s, 1H), 9.15 (s, 1H), 8.78 (s, 1H), 8.50-8.47 (m, 3H), 8.15 (s, 2H), 3.35 -3.29(m,1H)1.68(d, J =6.4Hz,2H).

Synthesis of compound of formula III

-7-基)脲之合成 Example 16. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(cyclopropyl ( (Methyl)amino)imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(200mg，656.76μmol)之THF(4mL)溶液中，添加N-甲基環丙胺(141.3mg，1.31mmol，364.05uL，HCl)。懸浮液於室溫攪拌3小時。LCMS顯示大部分反應物被耗盡。去除溶劑，得到2-氯-8-[環丙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-羧酸乙酯(186mg，631.07μmol，96.09%產率)，為黃色固體。LC-MS：m/z 295.1[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

7-carboxylic acid ethyl ester 7c (200mg, 656.76 μ mol) of THF (4mL) was added N- methyl cyclopropylamine (141.3mg, 1.31mmol, 364.05uL, HCl ). The suspension was stirred at room temperature for 3 hours. LCMS showed that most of the reactants were consumed. Removal of the solvent yields 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]

7-carboxylate (186mg, 631.07 μ mol, 96.09 % yield) as a yellow solid. LC-MS: m/z 295.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13a(186mg，631.07μmol)之乙醇(4mL)溶液中，添加氫氧化鋰一水合物(1M，1.89mL)。於室溫攪拌混合物10小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。過濾懸浮液，將濾餅真空乾燥，得到2-氯-8-[環丙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-羧酸(168mg，629.96μmol，99.82%產率)，為白色固體。 0℃, at 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]ta

7-carboxylic acid ethyl ester 13a (186mg, 631.07 μ mol) with ethanol (4mL) was added lithium hydroxide monohydrate (1M, 1.89mL). The mixture was stirred at room temperature for 10 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The suspension was filtered and the filter cake was vacuum dried to obtain 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]ta

7-carboxylic acid (168mg, 629.96 μ mol, 99.82 % yield) as a white solid.

Step 3

-7-羧酸14a(157mg，588.71μmol)之1,4-二

烷(7mL)溶液中，添加三乙胺(297.9mg，2.94mmol， 410.27uL)與[疊氮(苯氧)磷醯基]氧苯(194.42mg，706.46μmol，153.08uL)。攪拌混合物30分鐘。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(149.70mg，765.33μmol)，於100℃攪拌反應混合物2小時。濃縮混合物得到殘留物，利用製備型TLC(二氯甲烷：甲醇=20：1)將其純化，得到1-[2-氯-8-[環丙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(42.7mg，92.97μmol，15.79%產率)，為白色固體。LC-MS：m/z 459[M+H]⁺。¹H NMR(DMSO-d ₆,400MHz)δ 9.73(s,1H),8.55-8.47(m,4H),8.23(d,J=6.4Hz,1H),8.15(s,2H),3.22-3.17(m,4H),0.67-0.51(m,4H)。 At room temperature, in 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]

-7-carboxylic acid 14a (157mg, 588.71 μ mol) of 1,4-bis

Dioxane (7 mL) was added triethylamine (297.9mg, 2.94mmol, 410.27uL) and oxybenzone [azide (phenoxy) phosphoryl acyl] (194.42mg, 706.46 μ mol, 153.08uL). The mixture was stirred for 30 minutes. Was then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine 10a (149.70mg, 765.33 μ mol) , stirring the reaction mixture for 2 hours at 100 deg.] C. The mixture was concentrated to obtain the residue, which was purified by preparative TLC (dichloromethane: methanol=20:1) to obtain 1-[2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1 ,2-b]

7-yl] -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (42.7mg, 92.97 μ mol, 15.79 % yield) as a white solid. LC-MS: m/z 459 [M+H] ⁺ . ¹ H NMR(DMSO- d ₆ ,400MHz) δ 9.73(s,1H),8.55-8.47(m,4H),8.23(d, J =6.4Hz,1H),8.15(s,2H),3.22-3.17 (m, 4H), 0.67-0.51 (m, 4H).

-7-基)脲之合成 Example 17. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(dimethylamino) )Imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(300mg，985.13μmol)之THF(10mL)溶液中，添加N-甲基甲胺(2M，2.46mL)。該懸浮液於室溫攪拌1小時。TLC顯示大部分反應物被耗盡並出現一個主要斑點。濃縮混合物得到殘留物，利用矽膠管柱層析法以石油醚/乙酸乙酯(10/1)洗提將其純化，得到2-氯-8-(二甲 胺基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(250mg，930.41μmol，94.45%產率)，為黃色固體。LC-MS：m/z 269.1[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

7-carboxylic acid ethyl ester 7c (300mg, 985.13 μ mol) of THF (10mL) was added methyl N- methylamine (2M, 2.46mL). The suspension was stirred at room temperature for 1 hour. TLC showed that most of the reactants were consumed and a major spot appeared. The mixture was concentrated to obtain the residue, which was purified by elution with petroleum ether/ethyl acetate (10/1) by silica gel column chromatography to obtain 2-chloro-8-(dimethylamino)imidazo[1,2 -b] Da

7-carboxylate (250mg, 930.41 μ mol, 94.45 % yield) as a yellow solid. LC-MS: m/z 269.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13b(100mg，372.16μmol)之乙醇/水(5/5mL)溶液中，添加氫氧化鋰一水合物(1M，1.12mL)。於室溫攪拌混合物10小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。過濾懸浮液，將濾餅真空乾燥，得到2-氯-8-(二甲胺基)咪唑并[1,2-b]嗒

-7-羧酸(89.6mg，粗產物)，為黃色固體，直接用於下一步驟。 0°C. To 2-chloro-8-(dimethylamino)imidazo[1,2-b]ta

7-carboxylate 13b (100mg, 372.16 μ mol) of ethanol / water (5 / 5mL) was added lithium hydroxide monohydrate (1M, 1.12mL). The mixture was stirred at room temperature for 10 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The suspension was filtered and the filter cake was vacuum dried to obtain 2-chloro-8-(dimethylamino)imidazo[1,2-b]

-7-carboxylic acid (89.6 mg, crude product), as a yellow solid, was used directly in the next step.

Step 3

-7-羧酸14b(82mg，340.75μmol)之1,4-二

烷(7mL)溶液中，添加三乙胺(172.4mg，1.70mmol，237.47uL)與[疊氮(苯氧)磷醯基]氧苯(112.5mg，408.90μmol，88.61uL)。然後攪拌該混合物30分鐘，接著添加5-氯-6-(三唑-2-基)吡啶-3-胺(86.7mg，442.97μmol)，於100℃攪拌反應混合物3小時。濃縮混合物得到殘留物，利用製備型HPLC將其純化，得到1-[2-氯-8-(二甲胺基)咪唑并[1,2-b]嗒

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(7.0mg，16.16μmol，4.74%產率)，為白色固體。LC-MS：m/z 433[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆)δ 9.75(s,1H),8.57(d,J=1.6Hz,2H),8.46(d,J=1.6Hz,1H),8.32(s,1H),8.23(s,1H),8.15(s,2H),3.24(s,6H)。 At room temperature, in 2-chloro-8-(dimethylamino)imidazo[1,2-b]

-7-carboxylic acid 14b (82mg, 340.75 μ mol) of 1,4-bis

Dioxane (7 mL) was added triethylamine (172.4mg, 1.70mmol, 237.47uL) with [azide (phenoxy) phosphoryl acyl] oxy benzene (112.5mg, 408.90 μ mol, 88.61uL ). The mixture was then stirred for 30 minutes, followed by addition of 5-chloro-6- (triazol-2-yl) pyridin-3-amine (86.7mg, 442.97 μ mol), the reaction was stirred at 100 deg.] C for 3 hours. The mixture was concentrated to obtain a residue, which was purified by preparative HPLC to obtain 1-[2-chloro-8-(dimethylamino)imidazo[1,2-b]

7-yl] -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (7.0mg, 16.16 μ mol, 4.74 % yield) as a white solid. LC-MS: m/z 433 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO- d ₆ ) δ 9.75(s,1H), 8.57(d, J =1.6Hz,2H), 8.46(d, J =1.6Hz,1H), 8.32(s,1H), 8.23 (s, 1H), 8.15 (s, 2H), 3.24 (s, 6H).

-7-基)脲之合成 Example 18. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(N-morpholine) Base) imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(100.0mg，328.4μmol)之THF(3mL)混合物中，添加嗎啉(57.2mg，656.8μmol)。於25℃攪拌反應混合物隔夜。過濾分離固體，真空濃縮濾液。殘留物利用管柱層析法(以PE/EA=10/1洗提)純化，得到2-氯-8-(N-嗎啉基)-咪唑并[1,2-b]嗒

-7-羧酸乙酯(82mg，80.36%產率)，為黃色固體。LC-MS：m/z 311[M+H]⁺。 25℃, at 8-bromo-2-chloro-imidazo[1,2-b]

To a mixture of -7-carboxylic acid ethyl ester 7c (100.0 mg, 328.4 μ mol) in THF (3 mL) was added morpholine (57.2 mg, 656.8 μ mol). The reaction mixture was stirred at 25°C overnight. The solid was separated by filtration, and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-(N-morpholinyl)-imidazo[1,2-b]

Ethyl -7-carboxylate (82 mg, 80.36% yield), as a yellow solid. LC-MS: m/z 311 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13c(60mg，193.09μmol)之THF(6mL)與乙醇(1mL)溶液中，添加氫氧化鋰一水合物(81.0mg，1.93mmol)之水(1mL)溶液。於60℃攪拌混合物48小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取該混合物，乾燥有機相，濃縮，得到2-氯-8-(N-嗎啉基)-咪唑并[1,2-b]嗒

-7-羧酸(54.6mg，定量)，為白色固體，直接用於下一步驟。LC-MS：m/z 267[M+H]⁺。 To 2-chloro-8-(N-morpholinyl)-imidazo[1,2-b]ta

7-carboxylate 13c (60mg, 193.09 μ mol) of THF (6mL) and ethanol (1 mL) was added lithium hydroxide monohydrate (81.0mg, 1.93mmol) of water (1mL) was added. The mixture was stirred at 60°C for 48 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc, the organic phase was dried, and concentrated to give 2-chloro-8-(N-morpholinyl)-imidazo[1,2-b]

-7-carboxylic acid (54.6 mg, quantitative), as a white solid, was used directly in the next step. LC-MS: m/z 267 [M+H] ⁺ .

Step 3

烷(6mL)溶液中，添加疊氮膦酸二苯酯(59.4mg，229.23μmol，46.79μL)與三乙胺(96.6mg，955.13μmol，133.13μL)。所得懸浮液於室溫攪拌30分鐘。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(56.1mg，286.54μmol)，於100℃攪拌該混合物2小時。去除溶劑，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)與製備型HPLC純化，得到1-(2-氯-7-(N-嗎啉基)-吡唑并[1,5-a]嘧啶-6-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(18mg，19.83%產率)，為白色固體。LC-MS：m/z 475.0[M+H]⁺。¹H NMR(DMSO-d6,400MHz)：δ 9.89(brs,1H),8.58(d,J=2.4Hz,2H),8.50(s,1H),8.48(d,J=2.4Hz,1H),8.29(s,1H),8.15(s,2H),3.78(t,J=4.0Hz,4H),3.78(t,J=4.0Hz,4H)。 In 2-chloro-7-(N-morpholinyl)-pyrazolo[1,5-a]pyrimidine-6-carboxylic acid 14c (54mg, 191.03 μ mol) of 1,4-bis

In the alkane (6 mL) solution, add diphenyl azide phosphonate (59.4 mg, 229.23 μ mol, 46.79 μ L) and triethylamine (96.6 mg, 955.13 μ mol, 133.13 μ L). The resulting suspension was stirred at room temperature for 30 minutes. Was then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (56.1mg, 286.54 μ mol), and the mixture was stirred at 100 ℃ 2 hours. The solvent was removed, and the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) and preparative HPLC to obtain 1-(2-chloro-7-(N-morpholinyl)-pyrazole And [1,5-a]pyrimidin-6-yl)-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (18mg, 19.83% yield), white solid. LC-MS: m/z 475.0 [M+H] ⁺ . ¹ H NMR(DMSO- d6 ,400MHz): δ 9.89(brs,1H), 8.58(d, J =2.4Hz,2H), 8.50(s,1H), 8.48(d, J =2.4Hz,1H), 8.29 (s, 1H), 8.15 (s, 2H), 3.78 (t, J = 4.0 Hz, 4H), 3.78 (t, J = 4.0 Hz, 4H).

-7-基)脲之合成 Example 19. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-((2-methyl (Oxyethyl) (methyl) amino) imidazo[1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(90mg，0.3mmol)之THF(1mL)溶液中，添加2-甲氧-N-甲基-乙胺(131.7mg，1.48mmol)。於室溫攪拌所得混合物2小時。TLC顯示反應完全。濃縮反應混合物，利用快速矽膠層析 法(PE中0~10% EtOAc)純化，得到2-氯-8-[2-甲氧基乙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-羧酸乙酯(90mg，97%產率)，為白色固體。LC-MS：m/z 313.1[M+H]⁺。 At room temperature, in 8-bromo-2-chloro-imidazo[1,2- b ]

To a solution of ethyl-7-carboxylate 7c (90 mg, 0.3 mmol) in THF (1 mL) was added 2-methoxy-N-methyl-ethylamine (131.7 mg, 1.48 mmol). The resulting mixture was stirred at room temperature for 2 hours. TLC showed that the reaction was complete. The reaction mixture was concentrated and purified by flash silica gel chromatography (0-10% EtOAc in PE) to obtain 2-chloro-8-[2-methoxyethyl(methyl)amino]imidazo[1,2- b]

Ethyl -7-carboxylate (90 mg, 97% yield), a white solid. LC-MS: m/z 313.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13d(100mg，319.7μmol)與氫氧化鋰(153.2mg，6.4mmol)於THF(3mL)、水(2mL)與甲醇(2mL)中之混合物12小時。LC-MS顯示反應完全。減壓去除溶劑，使殘留物溶於水(10mL)中，以EA(5mLx2)萃取。其水相以1M HCl酸化至pH~6，以DCM(10mLx5)萃取，合併之有機層以無水Na₂SO₄乾燥，過濾及濃縮，得到2-氯-8-[2-甲氧基乙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-羧酸(70mg，77%產率)，為黃色固體，直接用於下一步驟。LC-MS：m/z 285.1[M+H]⁺。 Stir 2-chloro-8-[2-methoxyethyl(methyl)amino]imidazo[1,2-b]ta at 50℃

7-carboxylate 13d (100mg, 319.7 μ mol) and lithium hydroxide (153.2mg, 6.4mmol) in THF (3mL), a mixture of water (2mL) and methanol (2mL) in the 12 hours. LC-MS showed that the reaction was complete. The solvent was removed under reduced pressure, the residue was dissolved in water (10 mL), and extracted with EA (5 mL×2). The aqueous phase was acidified with 1M HCl to pH~6, extracted with DCM (10mLx5), the combined organic layer was dried with anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain 2-chloro-8-[2-methoxyethyl (Methyl)amino]imidazo[1,2- b ]ta

-7-carboxylic acid (70 mg, 77% yield), as a yellow solid, was used directly in the next step. LC-MS: m/z 285.1 [M+H] ⁺ .

Step 3

-7-羧酸14d(70mg，245.9μmol)之1,4-二

烷溶液中，添加[疊氮(苯基)磷醯基]苯(77.7mg，319.6μmol)與三乙胺(124.4mg，1.23mmol)。攪拌所得混合物30分鐘。添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(72.1mg，368.8μmol)，於100℃攪拌混合物2小時。LC-MS顯示反應完全，以乙酸乙酯(20mL)稀釋，以鹽水(10mLx2)洗滌。濃縮有機層，利用製備型TLC(以DCM：MeOH=20：1洗提)純化，得到1-[2-氯-8-[2-甲氧基乙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(14.8mg，13%產率)，為白色固體。LC-MS：m/z 477.0[M+H]⁺。¹H NMR(400MHz,DMSO-d₆)：δ 9.79(s,1 H),8.57(d,J=2.4Hz,1H),8.50(s,1 H),8.47(d,J=2.0Hz,1 H),8.42(s,1 H),8.25(s,1 H),8.15(s,2 H),3.90(t,J=5.2Hz,2 H),3.55(t,J=5.6Hz,2 H),3.19-3.18(m,6 H)。 At room temperature, in 2-chloro-8-[2-methoxyethyl(methyl)amino]imidazo[1,2- b ]

-7-carboxylic acid 14d (70mg, 245.9 μ mol) of 1,4-bis

Dioxane was added [azido (phenyl) phosphoryl acyl] benzene (77.7mg, 319.6 μ mol) and triethylamine (124.4mg, 1.23mmol). The resulting mixture was stirred for 30 minutes. Of 5-chloro-6- (triazol-2-yl) pyridin-3-amine 10a (72.1mg, 368.8 μ mol) , the mixture was stirred at 100 deg.] C for 2 hours. LC-MS showed that the reaction was complete, diluted with ethyl acetate (20 mL), and washed with brine (10 mL×2). The organic layer was concentrated and purified by preparative TLC (eluted with DCM:MeOH=20:1) to obtain 1-[2-chloro-8-[2-methoxyethyl(methyl)amino]imidazo[ 1,2- b ]

-7-yl]-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (14.8 mg, 13% yield) as a white solid. LC-MS: m/z 477.0 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO-d ₆ ): δ 9.79(s,1 H), 8.57(d, J =2.4Hz,1H), 8.50(s,1 H), 8.47(d, J =2.0Hz, 1 H), 8.42 (s, 1 H), 8.25 (s, 1 H), 8.15 (s, 2 H), 3.90 (t, J = 5.2 Hz, 2 H), 3.55 (t, J = 5.6 Hz, 2 H), 3.19-3.18 (m, 6 H).

Example 20. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(7-cyclopropyl-2-methyl- Synthesis of 2H-pyrazolo[4,3-b]pyridine-6-yl)urea

step 1

At room temperature, add 4-amino-1-methyl-1H-pyrazole-3-carboxylic acid methyl ester (2.0g, 12.9mmol) in DMF (120mL) solution, add di(tertiary butyl) dicarbonate ) (3.38 g, 15.44 mmol). The reaction mixture was heated to 60°C for 5 hours. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was poured into water and extracted twice with ethyl acetate. The combined organic phases were dried over Na ₂ SO _4, washed with brine, filtered, and concentrated. The obtained residue was purified by silica gel column chromatography with 0% to 50% ethyl acetate in petroleum ether to obtain 4-(tertiary butoxycarbonylamino)-1-methyl-pyrazole- Methyl 3-carboxylate (2.5 g, 75.98% yield). LC-MS: m/z 256[M+H] ⁺ .

Step 2

At room temperature, in 4-(tertiary butoxycarbonylamino)-1-methyl-pyrazole-3-carboxylic acid methyl ester (2.5g, 9.79mmol) in tetrahydrofuran (24mL), add hydroxide A solution of lithium (586.4 mg, 24.48 mmol) in water (5 mL), and the reaction mixture was stirred at room temperature for 4 hours. The starting material was completely consumed and the required quality was detected by LC-MS. It was concentrated and acidified with HCl (1N) to pH=3~4. The reaction mixture was then filtered, and the filter cake was washed with water. Then it was dried in vacuum to obtain 4-(tertiary butoxycarbonylamino)-1-methyl-pyrazole-3-carboxylic acid (2.3 g, 97.35% yield) as a white solid. LC-MS: m/z 242 [M+H] ⁺ .

Step 3

In 4-(tertiary butoxycarbonylamino)-1-methyl-pyrazole-3-carboxylic acid (2.3g, 9.53mmol) in acetonitrile (8mL) and tetrahydrofuran (8mL), add 1,1 '-Hydroxydiimidazole (2.32g, 14.30mmol). The resulting reaction mixture was stirred at room temperature for 2 hours and magnesium chloride (907.6 mg, 9.53 mmol) and triethylamine (2.89 g, 28.60 mmol, 3.99 mL) were added. The reaction mixture was stirred at room temperature overnight. After the reaction was completed, the reaction mixture was poured into water and extracted twice with ethyl acetate. The combined organic phases were dried over Na ₂ SO _4, washed with brine, filtered, and concentrated. The obtained residue was purified by silica gel column chromatography with 0% to 30% ethyl acetate in petroleum ether to obtain 3-[4-(tertiary butoxycarbonylamino)-1-methyl- Pyrazol-3-yl]-3-oxo-propionic acid ethyl ester (2.26 g, 76.14% yield). LC-MS: m/z 312 [M+H] ⁺ .

Step 4

At room temperature, add 3-[4-(tertiary butoxycarbonylamino)-1-methyl-pyrazol-3-yl]-3-oxo-propionic acid ethyl ester (1.0g, 3.21mmol) To the dichloromethane (15 mL) solution of the solution, N,N-dimethylformamide dimethyl acetal (3.06 g, 25.70 mmol, 3.44 mL) was added dropwise. The reaction mixture was stirred at room temperature overnight. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography with 0% to 10% methanol in dichloromethane to obtain 7-hydroxy-2-methyl-pyrazolo[4,3-b ] Ethyl pyridine-6-carboxylate (603 mg, 84.87% yield). LC-MS: m/z 222 [M+H] ⁺ .

Step 5

Heat a solution of 7-hydroxy-2-methyl-pyrazolo[4,3-b]pyridine-6-carboxylic acid ethyl ester (300mg, 1.36mmol) in phosphorus oxychloride (20.79g, 135.62mmol) to 80°C for 3 hours. LC-MS showed that the starting material was consumed and the required mass was detected. The reaction mixture was concentrated, and the residue was purified by silica gel column chromatography with 0% to 50% ethyl acetate in petroleum ether to obtain 7-chloro-2-methyl-pyrazolo[4,3-b ] Ethyl pyridine-6-carboxylate (300 mg, 92.30% yield). LC-MS: m/z 240 [M+H] ⁺ .

Step 6

At room temperature, add 7-chloro-2-methyl-pyrazolo[4,3-b]pyridine-6-carboxylic acid ethyl ester (300mg, 1.25mmol) in N,N-dimethylformamide ( 12mL) solution, add cyclopropylboronic acid (1.08g, 12.52mmol) and sodium carbonate (398.0mg, 3.76mmol). The reaction mixture was degassed and _{exchanged with N 2} twice. Dichloro bis [di - tert.butyl (4-dimethylaminophenyl) phosphine] palladium (177.3mg, 250.4 μ mol) to the mixture, the resultant reaction mixture was heated to 100 deg.] C overnight. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was poured into water and extracted twice with ethyl acetate. The combined organic phases were washed with brine, dried over Na ₂ SO _4, filtered and concentrated. The residue obtained was purified by silica gel column chromatography with 0% to 20% ethyl acetate in petroleum ether to obtain 7-cyclopropyl-2-methyl-pyrazolo[4,3-b] Ethyl pyridine-6-carboxylate (70 mg, 22.80% yield). LC-MS: m/z 246 [M+H] ⁺ .

Step 7

At 0 ℃, in 7-cyclopropyl-2-methyl-pyrazolo [4,3-b]pyridine-6-carboxylic acid ethyl ester (85mg, 346.6 μ mol) in tetrahydrofuran (6mL) solution, one by one A solution of lithium hydroxide (20.75 mg, 866.37 μ mol) in water 1 mL) was added dropwise. The resulting reaction mixture was stirred at room temperature for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. The reaction solution was acidified with HCl (1N) to pH=3~4. Then it was extracted twice with ethyl acetate. The combined organic phase was washed with brine, _{dried over Na 2} SO ₄ , filtered and concentrated to give 7-cyclopropyl-2-methyl-pyrazolo[4,3-b]pyridine-6-carboxylic acid (68mg, 90.33% yield), it was a yellow solid, and it was used directly in the next step without further purification. LC-MS: m/z 218 [M+H] ⁺ .

Step 8

烷(6mL)溶液中，添加疊氮膦酸二苯酯(103.5mg，0.376mmol)與三乙胺(158.6mg，1.567mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(91.6mg，0.47mmol)，加熱混合物至100℃ 2小時。將反應溶液傾入水中，以乙酸乙酯萃取兩次。該有機物以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(7-環丙基-2-甲基-2H-吡唑并[4,3-b]吡啶-6-基)脲(13.1mg，10.21%產率)，為白色固體。LC-MS：m/z 410[M+H]⁺。¹H NMR(DMSO-d ₆ ,400MHz)：δ 9.70(s,1 H),8.74(s,1 H),8.56(d,J=2.0Hz,1 H),8.51(s,1 H),8.47(d,J=2.0Hz,1 H),8.42(s,1 H),8.14(s,2 H),4.15(s,3 H),2.23-2.19(m,1 H),1.71-1.67(m,2 H),1.1-1.06(m,2 H)。 In 7-cyclopropyl-2-methyl-pyrazolo[4,3-b]pyridine-6-carboxylic acid (68mg, 0.3134mmol) in 1,4-bis

To the alkane (6 mL) solution, add diphenyl azide phosphonate (103.5 mg, 0.376 mmol) and triethylamine (158.6 mg, 1.567 mmol). The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (91.6 mg, 0.47 mmol) was added, and the mixture was heated to 100°C for 2 hours. The reaction solution was poured into water, and extracted twice with ethyl acetate. The organic matter was _{dried with Na 2} SO ₄ and concentrated. The residue obtained was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(5-chloro-6- (2H-1,2,3-Triazol-2-yl)pyridin-3-yl)-3-(7-cyclopropyl-2-methyl-2H-pyrazolo[4,3-b]pyridine -6-yl)urea (13.1 mg, 10.21% yield) as a white solid. LC-MS: m/z 410 [M+H] ⁺ . ¹ H NMR(DMSO- d ₆ ,400MHz): δ 9.70(s,1 H), 8.74(s,1 H), 8.56(d, J =2.0Hz,1 H), 8.51(s,1 H), 8.47(d, J =2.0Hz,1 H), 8.42(s, 1 H), 8.14(s, 2 H), 4.15(s, 3 H), 2.23-2.19(m, 1 H), 1.71-1.67 (m, 2 H), 1.1-1.06 (m, 2 H).

-7-基)脲之合成 Example 21. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(methyl(oxy Etidine-3-yl)amino)imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(150.0mg，492.6μmol)之THF(3mL)溶液中，添加氧雜環丁烷-3-胺(72.0mg，985.1μmol)。於室溫攪拌混 合物隔夜。反應完全後，過濾分離固體及濃縮濾液。殘留物利用管柱層析法(以PE/EA=10/1洗提)純化，得到2-氯-8-(氧雜環丁烷-3-基胺基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(140.0mg，95.8%產率)，為白色固體。LC-MS：m/z 297[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

To a THF (3 mL) solution of ethyl-7-carboxylate 7c (150.0 mg, 492.6 μ mol), oxetane-3-amine (72.0 mg, 985.1 μ mol) was added. The mixture was stirred at room temperature overnight. After the reaction was completed, the solid was separated by filtration and the filtrate was concentrated. The residue was purified by column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-(oxetan-3-ylamino)imidazo[1,2-b ]despair

Ethyl -7-carboxylate (140.0 mg, 95.8% yield), a white solid. LC-MS: m/z 297 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13e(140mg，471.84μmol)之DMF(3mL)溶液中，添加NaH(90.4mg，2.4mmol，60%純度)。將所得溶液加溫至室溫並攪拌0.5小時。然後逐滴添加碘甲烷(401.8mg，2.8mmol)至該混合物中。於室溫攪拌所得混合物2小時。以水猝滅反應混合物並以乙酸乙酯(70mL)萃取。合併之有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。殘留物利用快速管柱層析法純化，得到2-氯-8-[甲基(氧雜環丁烷-3-基)胺基]咪唑并[1,2-b]嗒

-7-羧酸乙酯(60.0mg，40.9%產率)，為無色油。LC-MS：m/z 311[M+H]⁺。 0℃, at 2-chloro-8-(oxetan-3-ylamino)imidazo[1,2-b]ta

7-carboxylate 13e (140mg, 471.84 μ mol) of DMF (3mL) was added NaH (90.4mg, 2.4mmol, 60% purity). The resulting solution was warmed to room temperature and stirred for 0.5 hour. Then methyl iodide (401.8 mg, 2.8 mmol) was added dropwise to the mixture. The resulting mixture was stirred at room temperature for 2 hours. The reaction mixture was quenched with water and extracted with ethyl acetate (70 mL). The combined organic phases were washed with brine (100 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The residue was purified by flash column chromatography to obtain 2-chloro-8-[methyl(oxetan-3-yl)amino]imidazo[1,2-b]ta

Ethyl -7-carboxylate (60.0 mg, 40.9% yield) as a colorless oil. LC-MS: m/z 311 [M+H] ⁺ .

Step 3

-7-羧酸乙酯13f(60.0mg，193.1μmol)之THF(2mL)與乙醇(1mL)溶液中，添加氫氧化鋰一水合物(81.0mg，1.9mmol)之水(1mL)溶液。於50℃攪拌混合物48小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取該混合物，合併之有機相以無水Na₂SO₄乾燥，過濾並真空濃縮，得到2-氯-8-[甲基(氧雜環丁烷-3-基)胺基]咪唑并[1,2-b]嗒

-7-羧酸(54.5mg，99.8%產率)，為白色固體，直接用於下一步驟。LC-MS：m/z 283[M+H]⁺。 To 2-chloro-8-[methyl(oxetan-3-yl)amino]imidazo[1,2-b]ta

7-carboxylic acid ethyl ester 13f (60.0mg, 193.1 μ mol) of THF (2mL) and ethanol (1 mL) was added lithium hydroxide monohydrate (81.0mg, 1.9mmol) of water (1mL) was added. The mixture was stirred at 50°C for 48 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc, the combined organic phase was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to give 2-chloro-8-[methyl(oxetan-3-yl)amino]imidazo[ 1,2-b]

-7-carboxylic acid (54.5 mg, 99.8% yield), as a white solid, was used directly in the next step. LC-MS: m/z 283[M+H] ⁺ .

Step 4

烷(6mL)溶液中，添加疊氮膦酸二苯酯(60.5mg，233.5μmol)與三乙胺(98.4mg，972.8μmol)。所得溶液於室溫攪拌30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(57.1mg，291.9μmol)，於100℃攪拌該混合物2小時。去除溶劑，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)與製備型HPLC純化，得到標題化合物1-[2-氯-7-[甲基(氧雜環丁烷-3-基)胺基]吡唑并[1,5-a]嘧啶-6-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(6mg，6.5%產率)。LC-MS：m/z 475.0[M+H]⁺。¹H NMR(DMSO-d ₆ ,400MHz)：δ 8.92(s,1H),8.72-8.69(m,2H),8.58(d,J=2.0Hz,1H),8.49(d,J=2.4Hz,1H),8.29(s,1H),8.16(s,2H),5.08(t,J=6.4Hz,1H),4.73(t,J=6.4Hz,2H),4.63(t,J=6.4Hz,2H),3.08(s,3H)。 In 2-chloro-7-[methyl(oxetan-3-yl)amino]pyrazolo[1,5-a]pyrimidine-6-carboxylic acid 14e (55.0 mg, 194.6 μ mol) 1,4-Tues

To the alkane (6 mL) solution, add diphenyl azide phosphonate (60.5 mg, 233.5 μ mol) and triethylamine (98.4 mg, 972.8 μ mol). The resulting solution was stirred at room temperature for 30 minutes and then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (57.1mg, 291.9 μ mol), and the mixture was stirred at 100 ℃ 2 hours. The solvent was removed, and the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) and preparative HPLC to obtain the title compound 1-[2-chloro-7-[methyl(oxetan Alkyl-3-yl)amino]pyrazolo[1,5-a]pyrimidin-6-yl]-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (6mg, 6.5% yield). LC-MS: m/z 475.0 [M+H] ⁺ . ¹ H NMR(DMSO- d ₆ ,400MHz): δ 8.92(s,1H),8.72-8.69(m,2H),8.58(d, J =2.0Hz,1H), 8.49(d, J =2.4Hz, 1H), 8.29(s, 1H), 8.16(s, 2H), 5.08(t, J =6.4Hz,1H), 4.73(t, J =6.4Hz,2H), 4.63(t, J =6.4Hz, 2H), 3.08(s, 3H).

-7-基)脲之合成 Example 22. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(3-chloro-8-cyclopropylimidazo [1,2-b]

-7-yl) urea synthesis

step 1

In the DMF (10mL) suspension of 1-(tertiary butoxycarbonylamino)imidazole-2-carboxylic acid ethyl ester 3 (1.75g, 6.86mmol), slowly add 1-chloropyrrolidine-2,5 -A solution of diketone (0.92 g, 6.86 mmol) in DMF (5 mL). The reaction mixture was stirred at room temperature for 6 hours. After the reaction was completed, saturated sodium bicarbonate was added to the reaction mixture and extracted with EtOAc (2×100 mL). The combined organic phases were washed with brine (50mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate 10:1) to obtain 1-((tertiary butoxycarbonyl)amino)-5-chloro-1H-imidazole-2-carboxylic acid Ethyl ester (1.0 g, 3.46 mmol, 50.4% yield) as a colorless oil. LC-MS: m/z 290 [M+H] ⁺ .

Step 2

At 0°C, in 1-((tertiary butoxycarbonyl)amino)-5-chloro-1H-imidazole-2-carboxylic acid ethyl ester 4 (1.0g, 3.45mmol) in THF (20mL) solution, one by one Potassium 2-methylpropane-2-oleate (1M, 10.35mL) was added dropwise for 5 minutes, anhydrous ethyl acetate (760.27g, 8.63mmol, 0.84mL) was added to the mixture, and it was stirred at 0°C for 15 minutes . The ice bath was removed, and the resulting mixture was stirred at room temperature for 2 hours. TLC showed the formation of spots, the reaction was cooled to 0°C and treated with 1.0N HCl via dropwise addition until the suspension was completely dissolved. The resulting solution was extracted with EtOAc (2×40 mL). The combined organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated to obtain (Z)-3-(1-((tertiary butoxycarbonyl)amino)-5-chloro- 1H-imidazol-2-yl)-3-hydroxyethyl acrylate (720 mg, crude product) as a yellow solid. LC-MS: m/z 332[M+H] ⁺ .

Step 3

-7-羧酸乙酯(380mg，1.57mmol，72.46%產率)，為白色固體。LC-MS：m/z 242[M+H]⁺。 In (Z)-3-(1-((tertiary butoxycarbonyl)amino)-5-chloro-1H-imidazol-2-yl)-3-hydroxyethyl acrylate 5e (720mg, 2.17mmol) To the DCM (20 mL) solution, 1,1-dimethoxy-N,N-dimethyl-methylamine (1.94 g, 16.28 mmol, 2.18 mL) was added. The reaction mixture was stirred at room temperature for 3 hours. After the reaction was complete, the mixture was concentrated to obtain a crude product, which was recrystallized from ethyl acetate to obtain 3-chloro-8-hydroxyimidazo[1,2-b]

Ethyl -7-carboxylate (380 mg, 1.57 mmol, 72.46% yield) as a white solid. LC-MS: m/z 242 [M+H] ⁺ .

Step 4

-7-羧酸乙酯6e(380mg，1.57mmol)之1,4-二

烷(6mL)攪拌溶液中，添加磷醯溴(1.35g，4.72mmol，0.48mL)。於110℃攪拌反應混合物2小時。反應完全後，冷卻反應混合物至室溫，將反應混合物傾入水中，以EtOAc(2×10mL)萃取。合併之有機相以鹽水(10mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物利用快速管柱層析法(石油醚/乙酸乙酯=7：1)純化，得到8-溴-3-氯咪唑并[1,2-b]嗒

-7-羧酸乙酯(220mg，722.43μmol，45.94%產率)，為黃色固體。LC-MS：m/z 304[M+H]⁺。 At room temperature, in 3-chloro-8-hydroxyimidazo[1,2-b]

-7-carboxylic acid ethyl ester 6e (380mg, 1.57mmol) of 1,4-bis

To the stirred solution of alkane (6 mL), phosphonium bromide (1.35 g, 4.72 mmol, 0.48 mL) was added. The reaction mixture was stirred at 110°C for 2 hours. After the reaction was completed, the reaction mixture was cooled to room temperature, the reaction mixture was poured into water, and extracted with EtOAc (2×10 mL). The combined organic phases were washed with brine (10 mL) was washed, dried over anhydrous Na ₂ SO _4, filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (petroleum ether/ethyl acetate=7:1) to obtain 8-bromo-3-chloroimidazo[1,2-b]

7-carboxylate (220mg, 722.43 μ mol, 45.94 % yield) as a yellow solid. LC-MS: m/z 304 [M+H] ⁺ .

Step 5

-7-羧酸乙酯7e(220mg，722.43μmol)、環丙基硼酸(62.1mg，722.43mmol)之甲苯(4mL)溶液中，添加三環己膦(40.5mg，144.49μmol)、磷酸三鉀(383.4mg，1.81mmol)與二乙醯氧鈀(16.2mg，72.24μmol)。於100℃攪拌懸浮液4小時。TLC(石油醚：乙酸乙酯=5：1)顯示大部分起始原料被消耗並存在一個主要斑點。通過矽藻土墊過濾反應混合物。濾液以水(5mL)稀釋並以乙酸乙酯(10mL)萃取。有機相以鹽水洗滌，以Na₂SO₄乾燥，濃縮得到殘留物，利用矽膠管柱層析法以石油醚/乙酸乙酯(8/1)洗提將其純化，得到3-氯-8-環丙基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(136mg，511.86μmol，70.85%產率)，為黃色固體。LC-MS：m/z 266[M+H]⁺。 On 8-bromo-3-chloroimidazo[1,2-b]

-7-carboxylic acid ethyl ester 7e (220 mg, 722.43 μ mol), cyclopropylboronic acid (62.1 mg, 722.43 mmol) in toluene (4 mL) solution, add tricyclohexylphosphine (40.5 mg, 144.49 μ mol), phosphoric acid tribasic (383.4mg, 1.81mmol) and acetyl two oxygen palladium (16.2mg, 72.24 μ mol). The suspension was stirred at 100°C for 4 hours. TLC (petroleum ether: ethyl acetate = 5:1) showed that most of the starting material was consumed and a major spot was present. The reaction mixture was filtered through a pad of Celite. The filtrate was diluted with water (5 mL) and extracted with ethyl acetate (10 mL). The organic phase was washed with brine, _{dried with Na 2} SO ₄ and concentrated to obtain a residue, which was purified by silica gel column chromatography with petroleum ether/ethyl acetate (8/1) to obtain 3-chloro-8- Cyclopropyl-imidazo[1,2-b]ta

Ethyl -7-carboxylate (136 mg, 511.86 μ mol, 70.85% yield), as a yellow solid. LC-MS: m/z 266 [M+H] ⁺ .

Step 6

-7-羧酸乙酯8i(136mg，511.86μmol)之乙醇/水(5/5mL)溶液中，添加氫氧化鋰一水合物(1M，2.55mL)。所得反應混合物於室溫攪拌10小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。過濾懸浮液，將濾餅真空乾燥，得到3-氯-8-環丙基咪唑并[1,2-b]嗒

-7-羧酸(110.0mg，粗產物)，為黃色固體，直接用於下一步驟。 At 0℃, in 3-chloro-8-cyclopropyl-imidazo[1,2-b]

7-carboxylate 8i (136mg, 511.86 μ mol) of ethanol / water (5 / 5mL) was added lithium hydroxide monohydrate (1M, 2.55mL). The resulting reaction mixture was stirred at room temperature for 10 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The suspension was filtered and the filter cake was vacuum dried to obtain 3-chloro-8-cyclopropylimidazo[1,2-b]

-7-carboxylic acid (110.0 mg, crude product), as a yellow solid, was used directly in the next step.

Step 7

-7-羧酸9k(110mg，462.88μmol)之1,4-二

烷(4mL)溶液中，添加三乙胺(233.8mg，2.31mmol，321.20uL)與疊氮膦酸二苯酯(165.6mg，601.75μmol，130.40uL)，攪拌混合物30分鐘。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(90.5mg，462.88μmol)，於100℃攪拌反應混合物3小時。濃縮混合物得到殘留物，利用製備型HPLC將其純化，得到1-(3-氯-8-環丙基-咪唑并[1,2-b]嗒

-7-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(35.4mg，82.28μmol，17.78%產率)，為白色固體。LC-MS：m/z 430[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆)δ 9.75(s,1H),9.00(s,1H),8.78(s,1H),8.58(d,J=2.4Hz,1H),8.48(d,J=2.4Hz,1H),8.15(s,2H),7.75(s,1H),2.23-2.18(m,1H),1.85-1.81(m,2H),1.19-1.14(m,2H)。 At room temperature, in 3-chloro-8-cyclopropyl-imidazo[1,2-b]

-7-carboxylic acid 9k (110mg, 462.88 μ mol) of 1,4-bis

Dioxane (4mL) was added triethylamine (233.8mg, 2.31mmol, 321.20uL) with an azide diphenyl phosphonate (165.6mg, 601.75 μ mol, 130.40uL ), the mixture was stirred for 30 minutes. Was then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (90.5mg, 462.88 μ mol), the reaction was stirred at 100 deg.] C for 3 hours. The mixture was concentrated to obtain a residue, which was purified by preparative HPLC to obtain 1-(3-chloro-8-cyclopropyl-imidazo[1,2-b]

7-yl) -3- [5-chloro-6- (triazol-2-yl) -3-pyridinyl] urea (35.4mg, 82.28 μ mol, 17.78 % yield) as a white solid. LC-MS: m/z 430 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO- d ₆ ) δ 9.75(s,1H),9.00(s,1H),8.78(s,1H), 8.58(d, J =2.4Hz,1H), 8.48(d, J =2.4Hz, 1H), 8.15 (s, 2H), 7.75 (s, 1H), 2.23-2.18 (m, 1H), 1.85-1.81 (m, 2H), 1.19-1.14 (m, 2H).

-7-基)-3- (5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)脲之合成 Example 23. 1-(8-(azetidin-1-yl)-2-chloroimidazo[1,2-b]

Synthesis of -7-yl)-3- (5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)urea

step 1

-7-羧酸乙酯7c(100mg，0.33mmol)之THF(5mL)溶液中，添加氮雜環丁烷鹽酸鹽(92.6mg，0.99mmol)與三乙胺(100.2mg，0.99mmol)。所得反應混合物於室溫攪拌隔夜。耗盡起始原料並以LC-MS檢測到所需質量。濃縮反應混合物，所得殘留物利用快速管柱層析法(以於石油中0%至30%之乙酸乙酯洗提)純化，得到8-(氮雜環丁-1-基)-2-氯咪唑并[1,2-b]嗒

-7-羧酸乙酯(91mg，98.5%產率)，為白色固體。LC-MS：m/z 281[M+H]⁺。 At 0℃, in 8-bromo-2-chloroimidazo[1,2-b]

To a THF (5 mL) solution of ethyl-7-carboxylate 7c (100 mg, 0.33 mmol), azetidine hydrochloride (92.6 mg, 0.99 mmol) and triethylamine (100.2 mg, 0.99 mmol) were added. The resulting reaction mixture was stirred at room temperature overnight. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was concentrated, and the resulting residue was purified by flash column chromatography (eluted with 0% to 30% ethyl acetate in petroleum) to obtain 8-(azetidin-1-yl)-2-chloro Imidazo[1,2-b]

Ethyl -7-carboxylate (91 mg, 98.5% yield), a white solid. LC-MS: m/z 281 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13g(91mg，0.33mmol)之THF(5mL)與乙醇(5mL)溶液中，添加氫氧化鋰一水合物(136.5mg，3.25mmol)之水(3.2mL)溶液。於70℃攪拌反應混合物4小時。耗盡起始原料並以LC-MS檢測到所需質量。真空去除溶劑後，殘留物以HCl(1M)酸化至pH=3~4。然後以乙酸乙酯萃取，合併之有機相以Na₂SO₄乾燥，過濾及濃縮，得到8- (氮雜環丁-1-基)-2-氯咪唑并[1,2-b]嗒

-7-羧酸(81mg，98.9%產率)，為白色固體，不需進一步純化，直接用於下一步驟。LC-MS：m/z 253[M+H]⁺。 To 8-(azetidin-1-yl)-2-chloroimidazo[1,2-b]ta

To a solution of 13 g (91 mg, 0.33 mmol) of ethyl -7-carboxylate in THF (5 mL) and ethanol (5 mL), a solution of lithium hydroxide monohydrate (136.5 mg, 3.25 mmol) in water (3.2 mL) was added. The reaction mixture was stirred at 70°C for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. After the solvent was removed in vacuo, the residue was acidified with HCl (1M) to pH=3~4. Then it was extracted with ethyl acetate, and the combined organic phase was _{dried with Na 2} SO ₄ , filtered and concentrated to obtain 8-(azetidin-1-yl)-2-chloroimidazo[1,2-b]

-7-carboxylic acid (81 mg, 98.9% yield) was a white solid, and it was used directly in the next step without further purification. LC-MS: m/z 253 [M+H] ⁺ .

Step 3

-7-羧酸14f(81mg，0.32mmol)之1,4-二

烷(5mL)溶液中，添加疊氮膦酸二苯酯(115mg，0.42mmol)與三乙胺(162.6mg，1.61mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(87.7mg，0.45mmol)，加熱混合物至100℃ 2小時。將反應混合物傾入水中，以乙酸乙酯萃取兩次。合併之有機相以Na₂SO₄乾燥，濃縮，所得殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(8-(氮雜環丁-1-基)-2-氯咪唑并[1,2-b]嗒

-7-基)-3-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)脲(15.8mg)，為紫色固體。LC-MS：m/z 445[M+H]⁺。¹H NMR(DMSO-d ₆ ,400MHz)：δ 9.68(s,1 H),8.58(s,1 H),8.45(d,J=2.4Hz,1 H),8.19(s,1 H),8.15(s,1 H),8.13(s,2 H),7.91(s,1 H),4.62(t,J=7.6Hz,4 H),2.38-2.34(m,2 H)。 To 8-(azetidin-1-yl)-2-chloroimidazo[1,2-b]ta

-7-carboxylic acid 14f (81mg, 0.32mmol) of 1,4-bis

To the alkane (5 mL) solution, add diphenyl azide phosphonate (115 mg, 0.42 mmol) and triethylamine (162.6 mg, 1.61 mmol). The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (87.7 mg, 0.45 mmol) was added, and the mixture was heated to 100°C for 2 hours. The reaction mixture was poured into water and extracted twice with ethyl acetate. The combined organic phase was _{dried with Na 2} SO ₄ and concentrated. The resulting residue was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(8-(nitrogen) Etan-1-yl)-2-chloroimidazo[1,2-b]ta

-7-yl)-3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)urea (15.8 mg) as a purple solid. LC-MS: m/z 445 [M+H] ⁺ . ¹ H NMR(DMSO- d ₆ ,400MHz): δ 9.68(s,1 H), 8.58(s,1 H), 8.45(d, J =2.4Hz,1 H), 8.19(s,1 H), 8.15 (s, 1 H), 8.13 (s, 2 H), 7.91 (s, 1 H), 4.62 (t, J = 7.6 Hz, 4 H), 2.38-2.34 (m, 2 H).

-7-基)脲之合成 Example 24. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(pyrrolidine-1) -Based) imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(100.0mg，328.4μmol)之1,4-二

烷(6mL)溶液中，添加吡咯啶(23.4mg，328.4μmol)、碳酸銫(321.2mg，985.2μmol)、參(二亞苄基丙酮)二鈀(0)(30.1mg，32.84μmol)與4,5-雙(二苯膦基)-9,9-二甲基二苯并哌喃(38.0mg，65.68μmol)。N₂下，於100℃攪拌反應混合物2小時，冷卻反應混合物至室溫，真空濃縮得到殘留物，利用快速管柱層析法(以PE/EA=10/1洗提)將其純化，得到2-氯-8-吡咯啶-1-基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(85mg，87.8%產率)，為黃色固體。LC-MS：m/z 295[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

-7-carboxylic acid ethyl ester 7c (100.0mg, 328.4 μ mol) of 1,4-bis

Dioxane (6mL) was added pyrrolidine (23.4mg, 328.4 μ mol), cesium carbonate (321.2mg, 985.2 μ mol), ginseng (dibenzylideneacetone) dipalladium (0) (30.1mg, 32.84 μ mol ) and 4,5-bis (diphenylphosphino) -9,9-dimethyl xanthene (38.0mg, 65.68 μ mol). The _{reaction mixture was stirred at 100°C under N 2} for 2 hours, the reaction mixture was cooled to room temperature, and concentrated in vacuo to obtain a residue, which was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-pyrrolidin-1-yl-imidazo[1,2-b]ta

Ethyl -7-carboxylate (85 mg, 87.8% yield), as a yellow solid. LC-MS: m/z 295 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13h(85.0mg，288.4μmol)之EtOH(4mL)與H₂O(4mL)溶液中，添加氫氧化鋰一水合物(121.1mg，2.88mmol)。於70℃攪拌反應混合物16小時。以EtOAc(5mL)稀釋反應混合物並以HCl(1N)調至pH=6。水相以EtOAc(2×5mL)萃取，合併之有機相以鹽水(100mL)洗滌，以無水Na₂SO₄乾燥，過濾並真空濃縮，得到粗產物2-氯-8-吡咯啶-1- 基-咪唑并[1,2-b]嗒

-7-羧酸(70mg，91.0%產率)，為黃色固體，不需進一步純化直接用於下一步驟。 On 2-chloro-8-pyrrolidin-1-yl-imidazo[1,2-b]ta

To a 13h (85.0 mg, 288.4 μ mol) solution of ethyl -7-carboxylate in EtOH (4 mL) and H ₂ O (4 mL), lithium hydroxide monohydrate (121.1 mg, 2.88 mmol) was added. The reaction mixture was stirred at 70°C for 16 hours. The reaction mixture was diluted with EtOAc (5 mL) and adjusted to pH=6 with HCl (1N). The aqueous phase was extracted with EtOAc (2×5 mL), the combined organic phase was washed with brine (100 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo to obtain the crude product 2-chloro-8-pyrrolidin-1-yl -Imidazo[1,2-b]ta

-7-carboxylic acid (70 mg, 91.0% yield), as a yellow solid, was used directly in the next step without further purification.

Step 3

-7-羧酸14g(70mg，262.5μmol)之1,4-二

烷(4mL)溶液中，添加二苯基磷醯基疊氮化物(93.9mg，341.23μmol)、三乙胺(132.6mg，1.3mmol)。於室溫攪拌反應混合物30分鐘。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(66.8mg，341.2μmol)，於100℃攪拌該混合物2小時。冷卻反應混合物至室溫，去除溶劑，殘留物利用快速管柱層析法(以DCM：MeOH=20：1洗提)純化，得到1-(2-氯-8-吡咯啶-1-基-咪唑并[1,2-b]嗒

-7-基)-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(25mg，20.7%產率)，為白色固體。LC-MS：m/z 459.0[M+H]⁺。¹H NMR(DMSO-d ₆ ,400MHz)：δ 8.58(s,1H),8.45(d,J=2.4Hz,1H),8.17(s,1H),8.14(s,2H),7.94(s,1H),4.01(t,J=6.0Hz,4H),1.87(t,J=6.4Hz,4H)。 On 2-chloro-8-pyrrolidin-1-yl-imidazo[1,2-b]ta

-7-carboxylic acid 14g (70mg, 262.5 μ mol) of 1,4-bis

Dioxane (4mL) was added diphenylphosphoryl acyl azide (93.9mg, 341.23 μ mol), triethylamine (132.6mg, 1.3mmol). The reaction mixture was stirred at room temperature for 30 minutes. Was then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (66.8mg, 341.2 μ mol), and the mixture was stirred for 2 hours at 100 ℃. The reaction mixture was cooled to room temperature, the solvent was removed, and the residue was purified by flash column chromatography (eluted with DCM:MeOH=20:1) to obtain 1-(2-chloro-8-pyrrolidin-1-yl- Imidazo[1,2-b]

-7-yl)-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (25 mg, 20.7% yield) as a white solid. LC-MS: m/z 459.0 [M+H] ⁺ . ¹ H NMR(DMSO- d ₆ ,400MHz): δ 8.58(s,1H), 8.45(d, J =2.4Hz,1H), 8.17(s,1H), 8.14(s,2H),7.94(s, 1H), 4.01 (t, J = 6.0 Hz, 4H), 1.87 (t, J = 6.4 Hz, 4H).

-7-基)脲之合成 Example 25. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(ethyl(form Group) amino group) imidazo[1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(100mg，0.33mmol)之THF(4mL)溶液中，添加N-甲基乙胺(68.2mg，1.16mmol)。於室溫攪拌反應混合物隔夜。耗盡起始原料並以LC-MS檢測到所需質量。濃縮反應混合物，所得殘留物利用快速管柱層析法(以於石油中0%至30%之乙酸乙酯洗提)純化，得到2-氯-8-(乙基(甲基)胺基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(90mg，96.7%產率)，為淡黃色固體。LC-MS：m/z 283[M+H]⁺。 0℃, at 8-bromo-2-chloroimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 7c (100 mg, 0.33 mmol) in THF (4 mL), N-methylethylamine (68.2 mg, 1.16 mmol) was added. The reaction mixture was stirred at room temperature overnight. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was concentrated, and the resulting residue was purified by flash column chromatography (eluted with 0% to 30% ethyl acetate in petroleum) to obtain 2-chloro-8-(ethyl(methyl)amino) Imidazo[1,2-b]

Ethyl -7-carboxylate (90 mg, 96.7% yield), as a pale yellow solid. LC-MS: m/z 283[M+H] ⁺ .

Step 2

-7-羧酸乙酯13i(90mg，0.32mmol)之THF(5mL)與乙醇(5mL)溶液中，添加氫氧化鋰一水合物(67mg，1.60mmol)之水1.6mL)溶液。於70℃攪拌混合物4小時。耗盡起始原料並以LC-MS檢測到所需質量。真空去除溶劑得到殘留物，以HCl(1N)將其酸化至pH=3~4。所得混合物以乙酸乙酯萃取，合併之有機相以鹽水洗滌，以Na₂SO₄乾燥，過濾，濃縮，得到2-氯-8-(乙基(甲基)胺基)咪唑并[1,2-b]嗒

-7-羧酸(81mg，99.9%產率)，為淡黃色固體，直接用於下一步驟。LC-MS：m/z 255[M+H]⁺。 To 2-chloro-8-(ethyl(methyl)amino)imidazo[1,2-b]ta

To a solution of -7-carboxylic acid ethyl ester 13i (90 mg, 0.32 mmol) in THF (5 mL) and ethanol (5 mL), a solution of lithium hydroxide monohydrate (67 mg, 1.60 mmol) in water (1.6 mL) was added. The mixture was stirred at 70°C for 4 hours. The starting material was consumed and the required quality was detected by LC-MS. The solvent was removed in vacuo to obtain a residue, which was acidified to pH=3~4 with HCl (1N). The resulting mixture was extracted with ethyl acetate, and the combined organic phase was washed with brine, _{dried over Na 2} SO ₄ , filtered, and concentrated to give 2-chloro-8-(ethyl(methyl)amino)imidazo[1,2 -b] Da

-7-carboxylic acid (81 mg, 99.9% yield), as a pale yellow solid, was used directly in the next step. LC-MS: m/z 255 [M+H] ⁺ .

Step 3

-7-羧酸14h(81mg，0.3189mmol)之1,4-二

烷(5mL)溶液中，添加疊氮膦酸二苯酯(114.1mg，0.4146mmol)與三乙胺(161.3mg，1.594mmol)。於室溫攪拌所得混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(87mg，0.4465mmol)，此混合物於100℃加熱2小時。將混合物溶液傾入水中，以乙酸乙酯萃取兩次。合併之有機相以Na₂SO₄乾燥，過濾及濃縮，殘留物利用快速管柱層析法(以於石油中10%至80%之乙酸乙酯洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-(乙基 (甲基)胺基)咪唑并[1,2-b]嗒

-7-基)脲(7.4mg，5.2%)，為白色固體。LC-MS：m/z 447[M+H]⁺。¹H NMR(DMSO-d ₆ ,400MHz)：δ 9.77(s,1 H),8.56(d,J=2.4Hz,2 H),8.46(d,J=2.4Hz,1 H),8.42(s,1 H),8.23(s,1 H),8.14(s,2 H),3.70(q,J=7.2,14.4Hz,2 H),3.13(s,3 H),1.16(t,J=7.2Hz,3 H)。 To 2-chloro-8-(ethyl(methyl)amino)imidazo[1,2-b]ta

-7-carboxylic acid 14h (81mg, 0.3189mmol) of 1,4-bis

To the alkane (5 mL) solution, diphenyl azide phosphonate (114.1 mg, 0.4146 mmol) and triethylamine (161.3 mg, 1.594 mmol) were added. The resulting mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (87 mg, 0.4465 mmol) was added, and the mixture was heated at 100°C for 2 hours. The mixture solution was poured into water and extracted twice with ethyl acetate. The combined organic phase was _{dried with Na 2} SO ₄ , filtered and concentrated. The residue was purified by flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum) to obtain 1-(5-chloro -6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(ethyl(methyl)amino)imidazo[1, 2-b]

-7-yl)urea (7.4 mg, 5.2%) as a white solid. LC-MS: m/z 447 [M+H] ⁺ . ¹ H NMR(DMSO- d ₆ ,400MHz): δ 9.77(s,1 H), 8.56(d, J =2.4Hz,2 H), 8.46(d, J =2.4Hz,1 H), 8.42(s ,1 H), 8.23 (s, 1 H), 8.14 (s, 2 H), 3.70 (q, J = 7.2, 14.4 Hz, 2 H), 3.13 (s, 3 H), 1.16 (t, J = 7.2Hz, 3 H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 26. 1-(2-Chloro-8-(cyclopropyl(methyl)amino)imidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

step 1

-7-羧酸乙酯7c(200mg，656.76μmol)之THF(4mL)溶液中，添加N-甲基環丙胺(141.3mg，1.31mmol，364.0uL，HCl)。懸浮液於室溫攪拌3小時。LCMS顯示用盡大部分起始原料。去除溶劑，得到2-氯-8-[環丙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-羧酸乙酯(186mg，631.07μmol，96.09%產率)，為黃色固體。LC-MS：m/z 295.1[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

7-carboxylic acid ethyl ester 7c (200mg, 656.76 μ mol) of THF (4mL) was added N- methyl cyclopropylamine (141.3mg, 1.31mmol, 364.0uL, HCl ). The suspension was stirred at room temperature for 3 hours. LCMS showed most of the starting material was used up. Removal of the solvent yields 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]

7-carboxylate (186mg, 631.07 μ mol, 96.09 % yield) as a yellow solid. LC-MS: m/z 295.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13a(186mg，631.07μmol)之乙醇(4mL)溶液中，添加氫氧化鋰一水合物(1M，1.89mL)。於室溫攪拌混合物10小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。 過濾懸浮液，將濾餅真空乾燥，得到2-氯-8-[環丙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-羧酸(168mg，629.96μmol，99.82%產率)，為白色固體，直接用於下一步驟。 0℃, at 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]ta

7-carboxylic acid ethyl ester 13a (186mg, 631.07 μ mol) with ethanol (4mL) was added lithium hydroxide monohydrate (1M, 1.89mL). The mixture was stirred at room temperature for 10 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The suspension was filtered and the filter cake was vacuum dried to obtain 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]ta

7-carboxylic acid (168mg, 629.96 μ mol, 99.82 % yield) as a white solid which was used directly in the next step.

Step 3

-7-羧酸(168mg，629.96μmol)之1,4-二

烷(7mL)溶液中，添加三乙胺(318.7mg，3.15mmol，439.02uL)與疊氮膦酸二苯酯(208.0mg，755.95μmol，163.81uL)，所得反應混合物於室溫攪拌30分鐘。然後添加2-(三氟甲基)吡啶-4-胺(132.76mg，818.95μmol)，於100℃攪拌反應混合物2小時。濃縮反應混合物得到殘留物，利用製備型TLC(以二氯甲烷：甲醇=20：1洗提)將其純化，得到1-[2-氯-8-[環丙基(甲基)胺基]咪唑并[1,2-b]嗒

-7-基]-3-[2-(三氟甲基)-4-吡啶基]脲(13.4mg，31.47μmol，5.00%產率)，為白色固體。LC-MS：m/z 426[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆)δ 9.83(s,1H),8.53(d,J=5.6Hz,1H),8.48(s,2H),8.24(s,1H),8.04(d,J=2.0Hz,1H),7.64-7.62(m,1H),3.21(s,3H),3.18-3.14(m,1H),0.65-0.62(m,2H),0.51-0.49(m,2H)。 At room temperature, in 2-chloro-8-[cyclopropyl(methyl)amino]imidazo[1,2-b]

-7-carboxylic acid (168mg, 629.96 μ mol) of 1,4-bis

Dioxane (7 mL) was added triethylamine (318.7mg, 3.15mmol, 439.02uL) with an azide diphenyl phosphonate (208.0mg, 755.95 μ mol, 163.81uL ), the resulting reaction mixture was stirred at room temperature for 30 minutes . Was then added 2- (trifluoromethyl) pyridin-4-amine (132.76mg, 818.95 μ mol), stirring the reaction mixture for 2 hours at 100 deg.] C. The reaction mixture was concentrated to obtain the residue, which was purified by preparative TLC (eluted with dichloromethane: methanol = 20:1) to obtain 1-[2-chloro-8-[cyclopropyl(methyl)amino] Imidazo[1,2-b]

7-yl] -3- [2- (trifluoromethyl) -4-pyridinyl] urea (13.4mg, 31.47 μ mol, 5.00 % yield) as a white solid. LC-MS: m/z 426 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO- d ₆ ) δ 9.83(s,1H), 8.53(d, J =5.6Hz,1H), 8.48(s,2H), 8.24(s,1H), 8.04(d, J =2.0Hz, 1H), 7.64-7.62 (m, 1H), 3.21 (s, 3H), 3.18-3.14 (m, 1H), 0.65-0.62 (m, 2H), 0.51-0.49 (m, 2H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 27. 1-(2-Chloro-8-(isopropyl(methyl)amino)imidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

step 1

-7-羧酸乙酯7c(250.0mg，820.9μmol)之THF(5mL)溶液中，添加N-甲基丙-2-胺(600.4mg，8.21mmol)。所得反應混合物於室溫攪拌隔夜。反應完全後，過濾分離固體並真空濃縮濾液。殘留物利用管柱層析法(以PE/EA=10/1洗提)純化，得到2-氯-8-(異丙基(甲基)胺基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(82mg，80.36%產率)，為黃色固體。LC-MS：m/z 297.1[M+H]⁺。 On 8-bromo-2-chloro-imidazo[1,2-b]

7-carboxylic acid ethyl ester 7c (250.0mg, 820.9 μ mol) of THF (5mL) was added N- methyl-2-amine (600.4mg, 8.21mmol). The resulting reaction mixture was stirred at room temperature overnight. After the reaction was complete, the solid was separated by filtration and the filtrate was concentrated in vacuo. The residue was purified by column chromatography (eluted with PE/EA=10/1) to obtain 2-chloro-8-(isopropyl(methyl)amino)imidazo[1,2-b]ta

Ethyl -7-carboxylate (82 mg, 80.36% yield), as a yellow solid. LC-MS: m/z 297.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯13j(210.0mg，707.7μmol)之THF(4mL)與H₂O(4mL)液中，添加氫氧化鋰一水合物(297mg，7.08mmol)。60℃攪拌反應混合物48小時。去除溶劑後，以HCl(2N)酸化殘留物至pH=3~4。以EtOAc萃取所得混合物，合併之有機相以Na₂SO₄乾燥，過濾，濃縮，得到2-氯-8-(異丙基(甲基)胺基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(171.0mg，定量)，為白色固體，直接用於下一步驟。LC-MS：m/z 297.1[M+H]⁺。 To 2-chloro-8-(isopropyl(methyl)amino)imidazo[1,2-b]ta

7-carboxylic acid ethyl ester 13j (210.0mg, 707.7 μ mol) of THF (4mL) and H ₂ O (4mL) was added lithium hydroxide monohydrate (297mg, 7.08mmol). The reaction mixture was stirred at 60°C for 48 hours. After removing the solvent, acidify the residue with HCl (2N) to pH=3~4. The resulting mixture was extracted with EtOAc, and the combined organic phase was _{dried over Na 2} SO ₄ , filtered, and concentrated to give 2-chloro-8-(isopropyl(methyl)amino)imidazo[1,2-b]a

Ethyl -7-carboxylate (171.0 mg, quantitative), a white solid, was used directly in the next step. LC-MS: m/z 297.1 [M+H] ⁺ .

Step 3

-7-羧酸14i(171mg，636.4μmol)之1,4-二

烷(4mL)溶液中，添加疊氮膦酸二苯酯(227.7mg，229.23μ mol，827.3uL)與三乙胺(321.4mg，3.18mmol)。所得懸浮液於室溫攪拌30分鐘。然後添加2-(三氟甲基)吡啶-4-胺(103.2mg，636.40μmol)，所得反應混合物於100℃攪拌2小時。真空去除溶劑得到殘留物，利用快速管柱層析法(以DCM：MeOH=30：1洗提)與製備型HPLC將其純化，得到1-(2-氯-8-(異丙基(甲基)胺基)咪唑并[1,2-b]嗒

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲(46.0mg，16.9%產率)，為白色固體。LC-MS：m/z 428.0[M+H]⁺。¹H NMR(400MHz,DMSO-d ₆ )：δ 9.88(brs,1H),8.53(d,J=5.6Hz,1H),8.42-8.40(m,2H),8.24-8.23(m,1H),8.06-8.05(m,1H),7.65-7.63(m,1H),4.52-4.47(m,1H),2.95(s,3H),1.22(d,J=6.4Hz,6H)。 To 2-chloro-8-(isopropyl(methyl)amino)imidazo[1,2-b]ta

-7-carboxylic acid 14i (171mg, 636.4 μ mol) of 1,4-bis

Dioxane (4mL) was added azide diphenyl phosphonate (227.7mg, 229.23 μ mol, 827.3uL ) and triethylamine (321.4mg, 3.18mmol). The resulting suspension was stirred at room temperature for 30 minutes. Was then added 2- (trifluoromethyl) pyridin-4-amine (103.2mg, 636.40 μ mol), the resulting reaction mixture was stirred for 2 hours at 100 ℃. The solvent was removed in vacuo to obtain the residue, which was purified by flash column chromatography (eluted with DCM:MeOH=30:1) and preparative HPLC to obtain 1-(2-chloro-8-(isopropyl(formyl) Group) amino group) imidazo[1,2-b]

-7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea (46.0 mg, 16.9% yield) as a white solid. LC-MS: m/z 428.0 [M+H] ⁺ . ¹ H NMR (400MHz, DMSO- d ₆ ): δ 9.88 (brs, 1H), 8.53 (d, J = 5.6 Hz, 1H), 8.42-8.40 (m, 2H), 8.24-8.23 (m, 1H), 8.06-8.05 (m, 1H), 7.65-7.63 (m, 1H), 4.52-4.47 (m, 1H), 2.95 (s, 3H), 1.22 (d, J = 6.4 Hz, 6H).

-7-基)脲之合成 Example 28. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(cyclopropyl ( 2-Methoxyethyl)amino)imidazo[1,2-b]ta

-7-yl) urea synthesis

By using N- (2-methoxyethyl)cyclopropylamine instead of oxetane-3-amine as the starting material, Example 28 was prepared in a procedure similar to that described in Preparation Example 21. LC-MS: m/z 503.0 [M+H] ⁺ . ¹ H NMR(400MHz,DMSO- d ₆ ): δ 10.06(s,1H),8.61(s,1H),8.56(d, J =2.4Hz,1H), 8.48(d, J =2.0Hz,1H) ,8.40(s,1H),8.25(s,1H),8.15(s,2H),3.90(t, J =5.6Hz,2H),3.52(t, J =5.6Hz,2H),3.33-3.31( m, 1H), 3.14 (s, 3H), 0.68-0.66 (m, 2H), 0.51-0.49 (m, 2H).

-7-基)脲之合成 Example 29. ( S )-1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-( 3-methyl-N-morpholinyl) imidazo[1,2-b]

-7-yl) urea synthesis

By using (3S)-3-methylmorpholine instead of oxetane-3-amine as the starting material, Example 29 was prepared in a similar procedure to that described in Preparation Example 21. LC-MS: m/z 489.0[M+H] ⁺ _; ¹ H NMR (400MHz, DMSO- d6 ): δ 10.29(brs, 1H), 9.16(s, 1H), 8.73(s, 1H), 8.57( d, J =2.4Hz,1H),8.51(d, J =2.4Hz,1H),8.35(s,1H),8.16(s,2H),4.16-4.12(m,1H),3.90-3.79(m , 3H), 3.49-3.43 (m, 2H), 3.09-3.06 (m, 1H), 0.86-0.85 (m, 3H).

-7-基)脲之合成 Example 30. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-((2S,6R )-2,6-Dimethyl-N-morpholino)imidazo[1,2-b]

-7-yl) urea synthesis

By using ( 2S,6R )-2,6-dimethylmorpholine instead of oxetane-3-amine as the starting material, Example 30 was prepared in a similar procedure to that described in Preparation Example 21. LCMS m/z 503[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d6 ) δ 9.85(s,1H), 8.58(d, J =2.4Hz,1H), 8.52(s,1H), 8.50 (s,1H),8.47(d, J =2.4Hz,1H),8.27(s,1H),8.78(s,1H),8.15(s,2H),3.82-3.78(m,4H),2.99- 2.93 (m, 2H), 1.11 (d, J = 2.4 Hz, 6H).

-7-基)脲之合成 Example 31. (S)-1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-( 3-(Methoxymethyl)-N-morpholinyl)imidazo[1,2-b]ta

-7-yl) urea synthesis

By using (3 S )-3-(methoxymethyl)morpholine instead of oxetane-3-amine as the starting material, Example 31 was prepared in a similar procedure to that described in Preparation Example 21. LC-MS(ESI) m/z 518.9[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 10.09(s,1H),8.78(s,1H),8.58-8.57(m,1H ), 8.50-8.49 (m, 2H), 8.31 (s, 1H), 8.15 (s, 1H), 8.04 (s, 1H), 4.39 (s, 1H), 3.91-3.84 (m, 2H), 3.79- 3.75 (m, 1H), 3.71-3.65 (m, 1H), 3.50-3.39 (m, 4H), 3.08 (s, 3H).

-7-基)脲之合成 Example 32. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(1,1- Dioxide-N-thiomorpholinyl) imidazo[1,2-b]ta

-7-yl) urea synthesis

烷1,1-二氧化物代替氧雜環丁烷-3-胺作為起始原料，以類似製備實施例21所述程序製備實施例32。LC-MS：m/z 523[M+H]⁺；¹H NMR(400MHz；DMSO-d6)：δ 10.02(br,1H),8.79(s,1H),8.79(s,1H),8.59(d,J=2.4Hz,1H),8.49(d,J=2.4Hz,1H),8.36(s,1H),8.16(s,2H),3.86(d,J=5.2Hz,4H),3.42(s,4H)。 By using 1,4-thiol

Alkyl 1,1-dioxide was used as the starting material instead of oxetane-3-amine, and Example 32 was prepared by a procedure similar to that described in Preparation Example 21. LC-MS: m/z 523[M+H] ⁺ ; ¹ H NMR (400MHz; DMSO- d6 ): δ 10.02 (br, 1H), 8.79 (s, 1H), 8.79 (s, 1H), 8.59 ( d,J=2.4Hz,1H),8.49(d, J =2.4Hz,1H),8.36(s,1H),8.16(s,2H),3.86(d, J =5.2Hz,4H),3.42( s,4H).

-7-基)脲之合成 Example 33. (S)-1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-( 3-Methoxypyrrolidin-1-yl)imidazo[1,2-b]ta

-7-yl) urea synthesis

By using ( 3S )-3-methoxypyrrolidine instead of oxetan-3-amine as the starting material, Example 33 was prepared in a similar procedure to that described in Preparation Example 21. LC-MS: m/z 489[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 9.79(br,1H), 8.59(s,1H), 8.45(d,J=2.0Hz, 2H), 8.19 (s, 1H), 8.14 (s, 2H), 7.95 (s, 1H), 4.16-4.01 (m, 5), 3.17 (s, 3H), 2.06-2.05 (m, 1), 1.94 -1.89(m,1).

-7-基)脲之合成 Example 34. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(methylthio) Imidazo[1,2-b]

-7-yl) urea synthesis

Example 34 was prepared in a similar procedure to that described in Preparation Example 21 by using sodium methyl mercaptan instead of oxetane-3-amine as the starting material. LC-MS: m/z 436.0[M+H] ⁺ ; ¹ HNMR (400MHz, DMSO- d6 ): δ 10.28 (s, 1H), 9.02 (s, 1H), 8.98 (s, 1H), 8.56 (d , J =2.4Hz,1H), 8.47(d, J =2.4Hz,1H), 8.41(s,1H), 8.16(s,2H), 2.82(s,3H).

-7-基)脲之合成 Example 35. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(cyclopropyl ( Ethyl)amino)imidazo[1,2-b]ta

-7-yl) urea synthesis

Example 35 was prepared in a similar procedure to that described in Preparation Example 21 by using N-ethylcyclopropylamine instead of oxetane-3-amine as the starting material. LC-MS: m/z 473[M+H] ⁺ ; ¹ H NMR (400MHz; DMSO- d ₆ ): δ 9.83(s, 1H), 8.67(s, 1H), 8.55(d, J = 2.4Hz ,1H),8.48(d, J =2.4Hz,2H),8.26(s,1H),8.15(s,2H),3.71(q, J =7.2Hz,2H),3.26-3.22(m,1H) , 1.12 (t, J = 7.2 Hz, 3H), 0.69-0.65 (m, 2H), 0.51-0.48 (m, 2H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 36. 1-(2-Chloro-8-(ethyl(2-methoxyethyl)amino)imidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

Example 36 was prepared in a similar procedure to that described in Preparation Example 21 by using N-ethyl-2-methoxy-ethylamine instead of oxetane-3-amine as the starting material. LC-MS: m/z 458.1[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 10.13(s,1H),8.76(s,1H),8.55(d, J =4.8Hz, 2H),8.27(s,1H),8.07(d, J =1.6Hz,1H),7.63-7.61(m,1H),3.67(t, J =5.6Hz,2H),3.55(q, J =7.2 Hz, 2H), 3.41 (t, J = 5.6 Hz, 2H), 3.12 (s, 3H), 1.01 (t, J = 7.2 Hz, 3H).

-7-基)脲之合成 Example 37. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(ethyl (2 -Methoxyethyl)amino)imidazo[1,2-b]ta

-7-yl) urea synthesis

Example 37 was prepared in a procedure similar to that described in Preparation Example 21 by using N-ethyl-2-methoxy-ethylamine instead of oxetane-3-amine as the starting material.

LC-MS: m/z 490.9[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ) δ 10.13 (s, 1H), 8.80 (s, 1H), 8.63 (s, 1H), 8.56 ( d, J =2.4Hz,1H),8.48(d, J =2.4Hz,1H),8.27(s,1H),8.15(s,2H),3.68(t, J =6.0Hz,2H),3.57( q, J = 7.2 Hz, 2H), 3.43 (t, J = 6.0 Hz, 2H), 3.15 (s, 3H), 1.03 (t, J = 7.2 Hz, 3H).

-7-基)脲之合成 Example 38. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(diethylamino) )Imidazo[1,2-b]ta

-7-yl) urea synthesis

Example 38 was prepared in a procedure similar to that described in Preparation Example 21 by using N-ethylethylamine instead of oxetane-3-amine as the starting material.

LC-MS: m/z 461.0[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.51(brs, 1H), 8.50(s, 1H), 8.78(s, 1H), 8.57 (d, J =2.4Hz,1H),8.48(d, J =2.4Hz,1H),8.26(s,1H),8.15(s,2H),3.55(q, J =7.2Hz,4H),1.05 (t, J =7.2Hz,6H).

-7-基)脲之合成 Example 39. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(N-morpholinyl)imidazo [1,2-b]

-7-yl) urea synthesis

Example 39 was prepared in a similar procedure to that described in Preparation Example 21 by using morpholine instead of oxetane-3-amine as the starting material.

LC-MS: m/z 441.1; ¹ H NMR (400MHz, DMSO- d ₆ ): δ 9.80 (brs, 1H), 8.59 (d, J = 2.4 Hz, 1H), 8.48 (d, J = 2.4 Hz, 2H), 8.39(s, 1H), 8.15(s, 2H), 8.09(d, J =1.2Hz,1H), 7.60(d, J =1.2Hz,1H), 3.78(t, J =4.8Hz, 4H), 3.69 (t, J = 4.8 Hz, 4H).

-7-基)脲之合成 Example 40. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(3-methoxy Azetidin-1-yl)imidazo[1,2-b]ta

-7-yl) urea synthesis

Example 40 was prepared in a procedure similar to that described in Preparation Example 21 by using 3-methoxyazetidine hydrochloride instead of oxetane-3-amine as the starting material. LC-MS: m/z 475.0[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 8.59 (s, 1H), 8.45 (d, J = 2.4 Hz, 1H), 8.19 (s, 1H), 8.14 (s, 2H), 7.96 (s, 1H), 4.81-4.77 (m, 2H), 4.41-4.39 (m, 2H), 4.34-4.30 (m, 1H), 3.25 (s, 3H) .

-7-基)脲之合成 Example 41. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(3-methoxy -3-Methylazetidin-1-yl)imidazo[1,2-b]ta

-7-yl) urea synthesis

By using 3-methoxy-3-methyl-azetidine hydrochloride instead of oxetane-3-amine as the starting material, Example 41 was prepared in a similar procedure to that described in Preparation Example 21. LC-MS: m/z 489.0[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 8.59 (s, 1H), 8.45 (d, J = 2.0 Hz, 1H), 8.19 (s, 1H), 8.14(s, 2H), 7.96(s, 1H), 4.51(d, J =10.0Hz, 2H), 4.42(d, J =9.6Hz, 2H), 3.21(s, 3H), 1.46( s,3H).

-1-基)咪唑并[1,2-b]嗒

-7-基)脲之合成 Example 42. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(piper

-1-yl)imidazo[1,2-b]ta

-7-yl) urea synthesis

-1-羧酸三級丁酯代替氧雜環丁烷-3-胺作為起始原料，以類似製備實施例21所述程序製備實施例42。LC-MS m/z 474[M+H]⁺；¹H NMR(400MHz,DMSO-d ₆)δ 10.13(s,1H),8.76(s,1H),8.59(t,J=2.0Hz,1H),8.47(d,J=2.0Hz,1H),8.43(s,1H),8.26(s,1H),8.14(s,2H),3.54(s,4H),2.91(s,4H),1.90(d,J=1.2Hz,1H)。 By using piperidine

Instead of oxetane-3-amine as the starting material, tert-butyl-1-carboxylate was used as the starting material, and Example 42 was prepared by a procedure similar to that of Preparation Example 21. LC-MS m/z 474[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 10.13(s,1H),8.76(s,1H),8.59(t, J =2.0Hz,1H ), 8.47(d, J = 2.0Hz, 1H), 8.43(s, 1H), 8.26(s, 1H), 8.14(s, 2H), 3.54(s, 4H), 2.91(s, 4H), 1.90 (d, J = 1.2Hz, 1H).

-1-基)咪唑并[1,2-b]嗒

-7-基)脲之合成 Example 43. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(4-methyl Piper

-1-yl)imidazo[1,2-b]ta

-7-yl) urea synthesis

代替氧雜環丁烷-3-胺作為起始原料，以類似製備實施例21所述程序製備實施例43。LCMS m/z 488[M+H]⁺；¹H NMR(400MHz,DMSO-d ₆)δ 10.14(s,1H),8.80(s,1H),8.59(s,1H),8.46(s,1H),8.38(s,1H),8.25(s,1H),8.14(s,2H),3.61(s,4H),2.22(s,4H),1.90(s,3H)。 1-methylpiperidine

Instead of oxetane-3-amine as the starting material, Example 43 was prepared by a procedure similar to that described in Preparation Example 21. LCMS m/z 488[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 10.14(s,1H),8.80(s,1H),8.59(s,1H),8.46(s,1H ), 8.38 (s, 1H), 8.25 (s, 1H), 8.14 (s, 2H), 3.61 (s, 4H), 2.22 (s, 4H), 1.90 (s, 3H).

-7-基)-3-(5-甲基吡啶-3-基)脲之合成 Example 44. 1-(2-Chloro-8-(dimethylamino)imidazo[1,2-b]

Synthesis of -7-yl)-3-(5-methylpyridin-3-yl)urea

Example 44 Example 44 was prepared in a procedure similar to that described in Preparation Example 21 by using N-methylmethylamine instead of oxetane-3-amine as the starting material. LC-MS(ESI+)[(M+H)+]: 406.0; ¹ H NMR(400MHz,DMSO- d6 ) δ 10.58(br s,1H),8.80(s,1H),8.23(s,1H), 8.20 (s, 1H), 7.63 (s, 1H), 3.23 (s, 6H).

-7-基)脲之合成 Example 45. 1-(2-Chloro-6-(trifluoromethyl)pyridin-4-yl)-3-(2-chloro-8-(ethyl(2-methoxyethyl)amino)imidazole And [1,2-b]

-7-yl) urea synthesis

By using N -ethyl-2-methoxy-ethylamine and 2-chloro-6-(trifluoromethyl)pyridine-4-aminomethane instead of oxetane-3-amine and 5-chloro-6 -(2H-1,2,3-triazol-2-yl)pyridin-3-amine was used as the starting material, and Example 45 was prepared by a procedure similar to that described in Preparation Example 21. LC-MS: m/z 492.0[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 10.26(s,1H), 8.65(d, J =6.4Hz,2H), 8.28(s, 1H),7.94(d, J =1.4Hz,1H),7.84(d, J =1.4Hz,1H), 3.71(t, J =5.6Hz,2H),3.58(q, J =7.2Hz,2H) ,3.42(t, J =5.6Hz,2H),3.13(s,3H),1.02(t, J =7.2Hz,3H).

-7-基)脲之合成 Example 46. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-isopropylimidazo[1,2 -b] Da

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7a(300mg，1.11mmol)之甲苯(4mL)溶液中，添加異丙烯基硼酸(95.4mg，1.11mmol)、三環己膦(62.3mg，0.222mmol)與磷酸鉀(589.5mg，2.78mmol)。將懸浮液脫氣並與N₂交換兩次。添加乙酸鈀(II)(24.9mg，0.111mmol)至該混合物中。於100℃攪拌反應混合物12小時。耗盡起始原料並以LC-MS檢測到所需質量。真空濃縮混合物，所得殘留物以快速管柱層析法(以PE/EA=10/1洗提)純化，得到8-異丙烯基咪唑并[1,2-b]嗒

-7-羧酸乙酯(100.5mg，39.13%產率)，為黃色固體。 To 8-bromoimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 7a (300mg, 1.11mmol) in toluene (4mL), add isopropenylboronic acid (95.4mg, 1.11mmol), tricyclohexylphosphine (62.3mg, 0.222mmol) and potassium phosphate ( 589.5 mg, 2.78 mmol). The suspension was degassed and _{exchanged with N 2} twice. Palladium(II) acetate (24.9 mg, 0.111 mmol) was added to the mixture. The reaction mixture was stirred at 100°C for 12 hours. The starting material was consumed and the required quality was detected by LC-MS. The mixture was concentrated in vacuo, and the resulting residue was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain 8-isopropenyl imidazo[1,2-b]

Ethyl -7-carboxylate (100.5 mg, 39.13% yield), as a yellow solid.

LC-MS: m/z 232.1 [M+H].

Step 2

-7-羧酸乙酯8j(100mg，0.432mmol)之甲醇(5mL)溶液中，添加氧化鉑(IV)(15mg，0.256mmol)。於室溫，氫氣氛圍下，攪拌反應混合物隔夜。將其過濾並真空濃縮。粗產物利用快速管柱層析法純化(以PE/EA=10/1洗提)，得到產物8-異丙基咪唑并[1,2-b]嗒

-7-羧酸乙酯(94mg，93.19%產率)，為白色固體。 To 8-isopropenylimidazo[1,2-b]ta

To a solution of ethyl -7-carboxylate 8j (100 mg, 0.432 mmol) in methanol (5 mL), platinum(IV) oxide (15 mg, 0.256 mmol) was added. The reaction mixture was stirred overnight at room temperature under a hydrogen atmosphere. It was filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (eluted with PE/EA=10/1) to obtain the product 8-isopropylimidazo[1,2-b]

Ethyl -7-carboxylate (94 mg, 93.19% yield), a white solid.

LC-MS: m/z 234.1 [M+H] ⁺ .

Step 3

-7-羧酸乙酯8k(92mg，0.394mmol)之THF(2mL)溶液中，添加氫氧化鋰一水合物(82.8mg，1.97mmol)之水(0.4mL)溶液。於室溫攪拌混合物5小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取該混合物，乾燥合併之有機相，濃縮，得到8-異丙基咪唑并[1,2-b]嗒

-7-羧酸(80mg，98.84%產率)，為黃色固體。 To 8-isopropylimidazo[1,2-b]

To a THF (2 mL) solution of ethyl -7-carboxylate 8k (92 mg, 0.394 mmol), a water (0.4 mL) solution of lithium hydroxide monohydrate (82.8 mg, 1.97 mmol) was added. The mixture was stirred at room temperature for 5 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc, the combined organic phases were dried and concentrated to give 8-isopropylimidazo[1,2-b]

-7-carboxylic acid (80 mg, 98.84% yield), as a yellow solid.

LC-MS: m/z 206.1 [M+H] ⁺ .

Step 4

-7-羧酸9l(80mg，0.390mmol)之1,4-二

烷(6mL)溶液中，添加疊氮膦酸二苯酯(128.74mg，0.468mmol)與三乙胺(196.87mg，1.95mmol)。所得溶液於室溫攪拌30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(76.3mg，0.389mmol)，於100℃攪拌該混合物2小時。真空去除溶劑後，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)純化，得到1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-(8-異丙基咪唑并[1,2-b]嗒

-7-基)脲(14.8mg，9.54%產率)，為白色固體。 To 8-isopropylimidazo[1,2-b]

-7-carboxylic acid 9l (80mg, 0.390mmol) of 1,4-bis

To the alkane (6 mL) solution, add diphenyl azide phosphonate (128.74 mg, 0.468 mmol) and triethylamine (196.87 mg, 1.95 mmol). The resulting solution was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (76.3 mg, 0.389 mmol) was added, and the mixture was stirred at 100°C for 2 hours. After the solvent was removed in vacuo, the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) to obtain 1-[5-chloro-6-(triazol-2-yl)-3-pyridine Yl]-3-(8-isopropylimidazo[1,2-b]

-7-yl)urea (14.8 mg, 9.54% yield) as a white solid.

LC-MS: m/z 398.1[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 9.67 (s, 1H), 8.81 (s, 1H), 8.59-8.57 (m, 2 H) ,8.47-8.46(m,1 H),8.18(d, J =1.2Hz,1H),8.15(s,2 H),7.70(d, J =1.2Hz,1H),3.52-3.47(m,1H ), 1.51(d, J = 6.8Hz, 6H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 47. 1-(8-Isopropylimidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

By using 2-(trifluoromethyl)pyridin-4-amine instead of 5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-amine as the starting material, Example 47 was prepared similarly to the procedure described in Preparation Example 46.

LC-MS: m/z 365.1[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 9.78 (s, 1H), 8.77 (s, 1H), 8.56-8.54 (m, 2 H) ,8.18(d, J =1.2Hz,1H),8.06-8.05(m,1 H),7.70(d, J =1.2Hz,1H),7.63-7.61(m,1 H),3.51-3.44(m ,1H), 1.50(d, J = 7.2Hz, 6H).

-7-基)脲之合成 Example 48. 1-(5-Chloro-6-methoxypyridin-3-yl)-3-(8-isopropylimidazo[1,2-b]

-7-yl) urea synthesis

By using 5-chloro-6-methoxy-pyridin-3-amine instead of 5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-amine as the starting material , Example 48 was prepared by a procedure similar to that described in Preparation Example 46.

LC-MS: m/z 361.1[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 9.14(s,1H), 8.62-8.59(m,2H), 8.15-8.13(m,3H ), 7.67-7.66 (m, 1H), 3.91 (s, 3H), 3.53-3.46 (m, 1H), 1.50 (d, J = 7.2 Hz, 6H).

-7-基)脲之合成 Example 49. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-ethylimidazo[1,2- b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯8e(150mg，0.591mmol)之甲醇(5mL)溶液中，添加Pd/C(71.8mg，591.29μmol)，將反應混合物脫氣，以H₂吹洗數次，H₂氛圍下，於室溫攪拌3小時。過濾該混合物，濾餅以乙酸乙酯洗滌。濃縮合併之有機層，得到8-乙基咪唑并[1,2-b]嗒

-7-羧酸乙酯(129mg，粗產物)，為黃色油，不需進一步純化即用於下一步驟。 On 2-chloro-8-ethyl-imidazo[1,2-b]

A solution of 7-carboxylate 8e (150mg, 0.591mmol) of methanol (5 mL), was added Pd / C (71.8mg, 591.29 μ mol), the reaction mixture was degassed, purged several times to H _2, H _{Under 2} atmosphere, stir at room temperature for 3 hours. The mixture was filtered and the filter cake was washed with ethyl acetate. Concentrate the combined organic layer to obtain 8-ethylimidazo[1,2-b]

Ethyl -7-carboxylate (129 mg, crude product), as a yellow oil, was used in the next step without further purification.

LC-MS: m/z 220.1 [M+H] ⁺ .

Step 2

-7-羧酸乙酯8e2(129mg，0.588mmol)之THF(3mL)溶液中，添加氫氧化鋰(74.1mg，1.77mmol)。於室溫攪拌該混合物12小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。過濾懸浮液，將濾餅真空乾燥，得到8-乙基咪唑并[1,2-b]嗒

-7-羧酸(112mg，粗產物)，為黃色固體。 At 0℃, at 8-ethylimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 8e2 (129 mg, 0.588 mmol) in THF (3 mL), lithium hydroxide (74.1 mg, 1.77 mmol) was added. The mixture was stirred at room temperature for 12 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The suspension was filtered and the filter cake was vacuum dried to obtain 8-ethylimidazo[1,2-b]

-7-carboxylic acid (112 mg, crude product) as a yellow solid.

LC-MS: m/z 192.1 [M+H] ⁺ .

Step 3

-7-羧酸9c2(147mg，0.769mmol)之1,4-二

烷(7mL)溶液中，添加N,N-二乙基乙胺(389.0mg，3.84mmol)與[疊氮(苯氧)磷酸基]氧苯(253.9mg，0.922mmol)。於室溫攪拌該混合物30分鐘。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(195.5mg，0.999mmol)，於100℃攪拌反應混合物2小時。TLC(二氯甲烷：甲醇=20：1，UV)顯示多數起始原料用盡。濃縮混合物得到殘留物，利用製備型TLC(二氯甲烷：甲醇=20：1)將其純化，得到1-[5-氯-6-(三唑 -2-基)-3-吡啶基]-3-(8-乙基咪唑并[1,2-b]嗒

-7-基)脲(45.4mg，15.38%產率)，為白色固體。 At room temperature, in 8-ethylimidazo[1,2-b]

-7-carboxylic acid 9c2 (147mg, 0.769mmol) of 1,4-bis

To the alkane (7 mL) solution, N,N-diethylethylamine (389.0 mg, 3.84 mmol) and [azide (phenoxy) phosphate] oxybenzene (253.9 mg, 0.922 mmol) were added. The mixture was stirred at room temperature for 30 minutes. Then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (195.5 mg, 0.999 mmol) was added, and the reaction mixture was stirred at 100°C for 2 hours. TLC (dichloromethane: methanol = 20:1, UV) showed that most of the starting materials were used up. The mixture was concentrated to obtain the residue, which was purified by preparative TLC (dichloromethane:methanol=20:1) to obtain 1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]- 3-(8-Ethylimidazo[1,2-b]ta

-7-yl)urea (45.4 mg, 15.38% yield) as a white solid.

LC-MS: m/z 384.0[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ) δ 9.74(s, 1H), 8.81(s, 1H), 8.78(s, 1H), 8.57( d, J =2.4Hz,1H),8.48(d, J =2.4Hz,1H),8.19-8.16(m,3H),7.68(d, J =2.4Hz,1H),2.99(q, J =7.2 Hz, 2H), 1.28 (t, J = 7.2 Hz, 1H).

-7-基)脲 Example 50. 1-[5-Chloro-6-(triazol-2-yl)-3-pyridyl]-3-(3-fluoro-8-isopropyl-imidazo[1,2-b] despair

-7-yl)urea

step 1

-7-羧酸乙酯7a(540mg，2.00mmol)之H₂O(2.5mL)與1,4-二

烷(10mL)混合溶劑之混合物中，添加磷酸鉀(1.27g，6.00mmol)、2-異丙烯基-4,4,5,5-四甲基-1,3,2-二

硼烷(403.2mg，2.40mmol)與[1,1'-雙(二苯膦基)二茂鐵]二氯鈀(II)(146.3mg，199.94μmol)。所得混合物以N₂脫氣，回流攪拌8小時，冷卻至室溫，通過矽膠過濾。固體濾餅以乙酸乙酯(20mL x 3)洗滌，將濾液真空濃縮。殘留物利用矽膠快速管柱層析法(以EtOAc/石油醚，0~50%，v/v洗提)純化，得到8-異丙烯基咪唑并[1,2-b]嗒

-7-羧酸乙酯(250mg，54.07%產率)，為白色固體。LCMS(ESI⁺)[(M+H)⁺]：231.8。 To 8-bromoimidazo[1,2-b]

-7-carboxylic acid ethyl ester 7a (540mg, 2.00mmol) in H ₂ O (2.5mL) and 1,4-bis

Add potassium phosphate (1.27g, 6.00mmol), 2-isopropenyl-4,4,5,5-tetramethyl-1,3,2-bis

Borane (403.2mg, 2.40mmol) and [1,1'-bis (diphenylphosphino) ferrocene] dichloropalladium (II) (146.3mg, 199.94 μ mol). The resulting mixture was N ₂ degassed and stirred under reflux for 8 hours, cooled to room temperature, filtered through silica gel. The solid filter cake was washed with ethyl acetate (20 mL x 3), and the filtrate was concentrated in vacuo. The residue was purified by silica gel flash column chromatography (eluted with EtOAc/petroleum ether, 0-50%, v/v) to obtain 8-isopropenylimidazo[1,2-b]ta

Ethyl -7-carboxylate (250 mg, 54.07% yield), a white solid. LCMS (ESI ⁺ ) [(M+H) ⁺ ]: 231.8.

Step 2

-7-羧酸乙酯8l(250mg，1.08mmol)與披鈀活性碳(0.1g，1.08mmol)於MeOH(5mL)中之混合物攪拌2小時，通過矽藻土墊過濾。以MeOH洗滌固體濾餅，真空濃縮濾液，得到粗產物8-異丙基咪唑并[1,2-b]嗒

-7-羧酸乙酯(180mg，71.38%產率)。LCMS(ESI⁺)[(M+H)⁺]：233.7。 Stir 8-isopropenylimidazo[1,2-b]ta at room temperature _{under H 2 atmosphere}

A mixture of 8 l (250 mg, 1.08 mmol) of ethyl -7-carboxylate and palladium activated carbon (0.1 g, 1.08 mmol) in MeOH (5 mL) was stirred for 2 hours and filtered through a pad of Celite. The solid filter cake was washed with MeOH, and the filtrate was concentrated in vacuo to obtain the crude product 8-isopropylimidazo[1,2-b]

Ethyl -7-carboxylate (180 mg, 71.38% yield). LCMS (ESI ⁺ ) [(M+H) ⁺ ]: 233.7.

Step 3

-7-羧酸乙酯8m(90mg，385.83μmol)之MeCN(5mL)溶液中，分數次添加Selecfluor(205.02mg，578.74μmol)。於此溫度攪拌混合物4小時，將其傾入H₂O(20mL)中，以乙酸乙酯(15mL x 2)萃取。合併之有機相以鹽水(20mL)洗滌，以無水Na₂SO₄乾燥，並予以過濾。真空濃縮濾液，殘留物利用矽膠快速管柱層析法(以於石油醚中10%至50%之乙酸乙酯洗提)純化，得到3-氟-8-異丙基-咪唑并[1,2-b]嗒

-7-羧酸乙酯(23mg，23.73%產率)，為白色固體。LCMS(ESI⁺)[(M+H)⁺]：251.6。 At 5℃~10℃, at 8-isopropylimidazo[1,2-b]

7-carboxylate 8m (90mg, 385.83 μ mol) of MeCN (5mL) solution, fractional added Selecfluor (205.02mg, 578.74 μ mol) . The mixture was stirred at this temperature for 4 hours, poured into H ₂ O (20 mL), and extracted with ethyl acetate (15 mL x 2). The combined organic phase was washed with brine (20 mL), dried over anhydrous Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica gel flash column chromatography (eluted with 10% to 50% ethyl acetate in petroleum ether) to obtain 3-fluoro-8-isopropyl-imidazo[1, 2-b]

Ethyl -7-carboxylate (23 mg, 23.73% yield), a white solid. LCMS (ESI ⁺ ) [(M+H) ⁺ ]: 251.6.

Step 4

-7-羧酸乙酯8n(23mg，91.54μmol)之THF(0.5mL)溶液中，添加氫氧化鋰一水合物(11.52mg，274.62μmol)之H₂O(0.2mL)溶液。於室溫攪拌該混合物隔夜並真空濃縮。殘留物以HCl水溶液(2M)酸化至pH 3~4。所得混合物以乙酸乙酯萃取(15mL x 2)，以無水硫酸鈉乾燥合併之有機相並予以過濾。真空濃縮濾液，得到粗產物3-氟-8-異丙基-咪唑并[1,2-b]嗒

-7-羧酸(18mg，88.10%產率)。LCMS(ESI⁺)[(M+H)⁺]：223.8。 On 3-fluoro-8-isopropyl-imidazo[1,2-b]

To a THF (0.5 mL) solution of ethyl-7-carboxylate 8n (23 mg, 91.54 μ mol), a solution of lithium hydroxide monohydrate (11.52 mg, 274.62 μ mol) in H ₂ O (0.2 mL) was added. The mixture was stirred at room temperature overnight and concentrated in vacuo. The residue was acidified to pH 3~4 with aqueous HCl (2M). The resulting mixture was extracted with ethyl acetate (15 mL x 2), and the combined organic phase was dried with anhydrous sodium sulfate and filtered. The filtrate was concentrated in vacuo to obtain the crude product 3-fluoro-8-isopropyl-imidazo[1,2-b]

-7-carboxylic acid (18 mg, 88.10% yield). LCMS (ESI ⁺ ) [(M+H) ⁺ ]: 223.8.

Step 5

-7-羧酸9m(18mg，80.64μmol)之1,4-二

烷(4mL)溶液中，添加疊氮膦酸二苯酯(31.4mg，120.97μmol，24.69uL)與TEA(24.5mg，241.93μmol，33.72uL)。於室溫攪拌所得混合物30分鐘，隨後添加5-氯-6-(三唑-2-基)吡啶-3-胺(15.8mg，80.64μmol)。於100℃攪拌該混合物隔夜，冷卻至室溫，將其傾入水中(10mL)並以乙酸乙酯(15mL x 2)萃取。合併之有機相以無水Na₂SO₄乾燥，並予以過濾。真空濃縮濾液，殘留物利用矽膠快速管柱層析法(以於石油醚中10%至80%之乙酸乙酯洗提)與製備型HPLC純化，得到1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-(3-氟-8-異丙基-咪唑并[1,2-b]嗒

-7-基)脲(4.0mg，11.93%產率)，為白色固體。LCMS(ESI⁺)[(M+H)⁺]：416.¹H NMR(400MHz,DMSO-d ₆)δ 8.97(s,1H),8.69(s,1H),8.59(d,J=2.4Hz,1H),8.47(d,J=2.4Hz,1H),8.16(s,2H),7.53(d,J=7.2Hz,1H),3.50(dt,J=13.6,6.8Hz,1H),1.51(d,J=6.8Hz,6H)。 On 3-fluoro-8-isopropyl-imidazo[1,2-b]

-7-carboxylic acid 9m (18mg, 80.64 μ mol) of 1,4-bis

To the alkane (4 mL) solution, add diphenyl azide phosphonate (31.4 mg, 120.97 μ mol, 24.69 uL) and TEA (24.5 mg, 241.93 μ mol, 33.72 uL). The resulting mixture was stirred at room temperature for 30 minutes, followed by the addition of 5-chloro-6- (triazol-2-yl) pyridin-3-amine (15.8mg, 80.64 μ mol). The mixture was stirred at 100°C overnight, cooled to room temperature, poured into water (10 mL) and extracted with ethyl acetate (15 mL x 2). The combined organic phase was dried with anhydrous Na ₂ SO ₄ and filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica gel flash column chromatography (eluted with 10% to 80% ethyl acetate in petroleum ether) and preparative HPLC to obtain 1-[5-chloro-6-(三(Azol-2-yl)-3-pyridyl]-3-(3-fluoro-8-isopropyl-imidazo[1,2-b]

-7-yl)urea (4.0 mg, 11.93% yield) as a white solid. ^{LCMS (ESI +) [(M} + H) +]:. 416 1 H NMR (400MHz, DMSO- d 6) δ 8.97 (s, 1H), 8.69 (s, 1H), 8.59 (d, J = 2.4Hz ,1H),8.47(d, J =2.4Hz,1H),8.16(s,2H),7.53(d, J =7.2Hz,1H), 3.50(dt, J =13.6,6.8Hz,1H),1.51 (d, J = 6.8Hz, 6H).

-7-基)脲之合成 Example 51. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(trifluoromethyl) )Imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7c(300mg，985.13μmol)，脫氣並再灌滿N₂三次，於50℃加熱18小時並經LCMS判斷反應完全，然後冷卻至室溫，以MTBE稀釋並通過矽藻土墊過濾。以MTBE洗滌該矽藻土墊，合併之有機層以鹽水洗滌，以Na₂SO₄乾燥。過濾後，蒸發溶劑，粗混合物利用矽膠管柱層析法純化，使用EA/PE=0：100-1：50為洗提液，得到2-氯-8-(三氟甲基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(80mg，272.45μmol，27.66%產率)，LC-MS：m/z 294[M+H]⁺。 To a stirred solution of cuprous chloride (195.1 mg, 1.97 mmol) and potassium tertiary butoxide 7c (220.7 mg, 1.97 mmol) was added 1,10-phenanthroline (355.06 mg, 1.97 mmol) in DMF (3.0 mL) The solution was degassed and filled with N ₂ three times. The dark red mixture was stirred at room temperature for 30 minutes, and then trimethyl (trifluoromethyl) silane (280.6 mg, 1.97 mmol, 313.03 uL) was slowly added. At room temperature, the resulting mixture was further stirred for 1 hour, and 8-bromo-2-chloro-imidazo[1,2-b] was added quickly

7-carboxylic acid ethyl ester 7c (300mg, 985.13 μ mol) , degassed and then filled with N ₂ three times, heated at 50 ℃ 18 h and judged complete by LCMS, cooled to room temperature, diluted with MTBE and by Filtered on a pad of diatomaceous earth. The celite pad was washed with MTBE, and the combined organic layer was washed with brine and dried _{with Na 2} SO _4. After filtration, the solvent was evaporated, and the crude mixture was purified by silica gel column chromatography, using EA/PE=0:100-1:50 as the eluent to obtain 2-chloro-8-(trifluoromethyl)imidazo[ 1,2-b]

7-carboxylate (80mg, 272.45 μ mol, 27.66 % yield), LC-MS: m / z 294 [M + H] +.

Step 2

-7-羧酸乙酯8o(80mg，272.45μmol)之THF(2.0mL)攪拌溶液中，添加LiOH(0.82mL,1M)，所得混合物於室溫攪拌隔夜，經LCMS判斷反應完全，然後添加3M鹽酸調至pH=2至3，並以EA(20mL)萃取，有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，得到粗產物2-氯-8-(三氟甲基)咪唑并[1,2-b]嗒

-7-羧酸(80mg，301.23μmol，110.56%產率)，粗產物直接用於下一步驟不需進一步純化。LC-MS：m/z 266[M+H]⁺。 To 2-chloro-8-(trifluoromethyl)imidazo[1,2-b]ta

7-carboxylate 8o (80mg, 272.45 μ mol) of THF (2.0mL) was stirred solution was added LiOH (0.82mL, 1M), and the resulting mixture was stirred at room temperature overnight, the reaction was judged complete by LCMS, and then add 3M hydrochloric acid was adjusted to pH=2 to 3, and extracted with EA (20 mL), the organic phase was washed with brine (5 mL), _{dried with Na 2} SO ₄ , filtered and concentrated in vacuo to obtain the crude product 2-chloro-8-(三Fluoromethyl)imidazo[1,2-b]ta

-7-carboxylic acid (80 mg, 301.23 μ mol, 110.56% yield), the crude product was used directly in the next step without further purification. LC-MS: m/z 266 [M+H] ⁺ .

Step 3

-7-羧酸9n(80mg，301.23μmol)之1,4-二

烷(2.0mL)攪拌溶液中，添加TEA(91.4mg，903.69μmol，125.96uL)與DPPA(124.4mg，451.85μmol，97.91uL)。於室溫攪拌反應混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺10a(88.4mg，451.85μmol)，於100℃攪拌反應混合物隔夜，經LCMS判斷反應完全，冷卻混合物至室溫並蒸發溶劑，粗混合物利用製備型HPLC純化，得到1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[2-氯-8-(三氟甲基)咪唑并[1,2-b]嗒

-7-基]脲(20mg，43.65μmol，14.49%產率)，LC-MS：m/z 459[M+H]⁺；¹H NMR(400MHz,DMSO-d6)δ 10.12(s,1H),9.26(s,1H),9.07(s,1H),8.61(s,1H),8.56(t,J=0.5Hz,1H),8.47(t,J=1.2Hz,1H),8.17(s,2H)。 To 2-chloro-8-(trifluoromethyl)imidazo[1,2-b]ta

-7-carboxylic acid 9n (80mg, 301.23 μ mol) of 1,4-bis

To the alkane (2.0 mL) stirring solution, TEA (91.4 mg, 903.69 μ mol, 125.96 uL) and DPPA (124.4 mg, 451.85 μ mol, 97.91 uL) were added. The reaction mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine 10a (88.4 mg, 451.85 μ mol) was added, and the reaction mixture was stirred at 100° C. overnight, and subjected to LCMS The reaction was judged to be complete, the mixture was cooled to room temperature and the solvent was evaporated. The crude mixture was purified by preparative HPLC to obtain 1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[2 -Chloro-8-(trifluoromethyl)imidazo[1,2-b]ta

7-yl] urea (20mg, 43.65 μ mol, 14.49 % yield), LC-MS: m / z 459 [M + H] +; 1 H NMR (400MHz, DMSO- d6) δ 10.12 (s, 1H ),9.26(s,1H),9.07(s,1H),8.61(s,1H),8.56(t, J =0.5Hz,1H),8.47(t, J =1.2Hz,1H),8.17(s ,2H).

-7-基)脲之合成 Example 52.1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(methoxymethyl Base) imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1

In a stirred solution of 2-(bromomethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (300mg, 1.36mmol) in MeOH (6mL), Add CH ₃ ONa (73.4 mg, 1.36 mmol). The reaction mixture was stirred at 25°C overnight, ice water (1 mL) was added, and extracted with EtOAc (40 mL). The organic phase was washed with brine (5 mL), _{dried over Na 2} SO ₄ , filtered and concentrated in vacuo to give the crude product 2- (Methoxymethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (240 mg, 1.40 mmol, 102.73% yield), a white solid.

Step 2

-7-羧酸乙酯(339.88mg，1.12mmol)之1,4-二

烷(5.0mL)、H₂O(0.5mL)攪拌溶液中，添加2-(甲氧甲基)-4,4,5,5-四甲基-1,3,2-二氧雜硼雜環戊烷(240mg，1.40mmol)、K₃PO₄(888.42mg，4.19mmol)，脫氣並再灌滿N₂三次，添加Pd(dppf)Cl₂．DCM(113.93mg，139.51μmol)並脫氣並再灌滿N₂三次，於90℃加熱混合物4小時並經LCMS判斷反應完全，然後冷卻，以EA稀釋並以鹽水洗滌，以Na₂SO₄乾燥。過濾並蒸發溶劑後，粗混合物利用矽膠管柱層析法純化，使用EA/PE=0：100-1：10作為洗提液，得到2-氯-8-(甲氧甲基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(80mg，296.64μmol，21.26%產率)，為白色固體。 On 8-bromo-2-chloro-imidazo[1,2-b]

-7-carboxylic acid ethyl ester (339.88mg, 1.12mmol) of 1,4-bis

Add 2-(methoxymethyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborole to the stirring solution of alkane (5.0mL) and H _{2 O (0.5mL)} Cyclopentane (240mg, 1.40mmol), K ₃ PO ₄ (888.42mg, 4.19mmol), degas and fill with N ₂ three times, add Pd(dppf)Cl ₂ . DCM (113.93mg, 139.51 μ mol) and degassed and then filled with N ₂ three times, the mixture was heated at 90 deg.] C for 4 hours and the reaction was judged complete by LCMS, then cooled, diluted with EA and washed with brine, Na ₂ SO ₄ to dry. After filtering and evaporating the solvent, the crude mixture was purified by silica gel column chromatography, using EA/PE=0:100-1:10 as the eluent to obtain 2-chloro-8-(methoxymethyl)imidazo[ 1,2-b]

7-carboxylate (80mg, 296.64 μ mol, 21.26 % yield) as a white solid.

LC-MS: m/z 270 [M+H] ⁺ .

Step 3

-7-羧酸乙酯8p(80mg，296.64μmol)之THF(4.0mL)攪拌溶液中，添加LiOH(0.5M，1.2ml)，於0℃攪拌2小時並經LCMS判斷反應完全，然後添加3M鹽酸調至pH=2至3並以EA(20mL)萃取，其有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，得到粗產物2-氯-8-(甲氧基甲基)咪唑并[1,2-b]嗒

-7-羧酸(60mg，248.31μmol，83.71%產率)，LC-MS：m/z 242[M+H]⁺，粗產物直接用於下一步驟不需進一步純化。 At 0℃, in 2-chloro-8-(methoxymethyl)imidazo[1,2-b]ta

7-carboxylate 8p (80mg, 296.64 μ mol) of THF (4.0mL) was stirred solution was added LiOH (0.5M, 1.2ml), stirred for 2 hours at 0 ℃ and judged complete by LCMS, and then add 3M hydrochloric acid was adjusted to pH=2 to 3 and extracted with EA (20mL). The organic phase was washed with brine (5mL), _{dried with Na 2} SO ₄ , filtered and concentrated in vacuo to obtain the crude product 2-chloro-8-(form (Oxymethyl) imidazo[1,2-b]

7-carboxylic acid (60mg, 248.31 μ mol, 83.71 % yield), LC-MS: m / z 242 [M + H] +, the crude product was directly used in the next step without further purification.

Step 4

-7-羧酸9o(60mg，248.31μmol)之1,4-二

烷(4mL)攪拌溶液中，添加TEA(75.4mg，744.94μmol，103.83uL)與DPPA(90.6mg，372.47μmol)。於室溫攪拌反應混合物2小時。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(58.3mg，297.98μmol)，於100℃攪拌反應混合物2小時並經LCMS判斷反應完全。冷卻混合物至室溫，蒸發溶劑，粗混合物利用製備型HPLC純化，得到1-[2-氯-8-(甲氧基甲基)咪唑并[1,2-b]嗒

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(14.0mg，32.24μmol),LC-MS：m/z 435[M+H]⁺；¹H NMR(400MHz,DMSO-d6)δ 9.05(s,1H),8.49(d,J=2.3Hz,1H),8.42(d,J=2.3Hz,1H),8.34(s,1H),8.09(s,2H),4.81(s,2H),3.32(s,3H)。 On 2-chloro-8-(methoxymethyl)imidazo[1,2-b]ta

-7-carboxylic acid 9o (60mg, 248.31 μ mol) of 1,4-bis

To the alkane (4 mL) stirring solution, TEA (75.4 mg, 744.94 μ mol, 103.83 uL) and DPPA (90.6 mg, 372.47 μ mol) were added. The reaction mixture was stirred at room temperature for 2 hours. Was then added 5-chloro-6- (triazol-2-yl) pyridin-3-amine (58.3mg, 297.98 μ mol), the reaction mixture was stirred for 2 hours at 100 deg.] C and the reaction was judged complete by LCMS. The mixture was cooled to room temperature, the solvent was evaporated, and the crude mixture was purified by preparative HPLC to obtain 1-[2-chloro-8-(methoxymethyl)imidazo[1,2-b]

-7-yl]-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (14.0 mg, 32.24 μ mol), LC-MS: m/z 435[M+ H] ⁺ ; ¹ H NMR(400MHz,DMSO- d6 ) δ 9.05(s,1H), 8.49(d, J =2.3Hz,1H), 8.42(d, J =2.3Hz,1H), 8.34(s, 1H), 8.09(s, 2H), 4.81(s, 2H), 3.32(s, 3H).

-7-基)脲之合成 Example 53. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(methoxymethyl)imidazo [1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯7a(1.5g，5.55mmol)之THF(20mL)溶液中，添加氫氧化鋰一水合物(699.2mg，16.66mmol)之水(10mL)溶液。於室溫攪拌混合物5小時。去除溶劑後，殘留物以HCl(2M)酸化至pH=3~4。以EtOAc萃取該混合物並乾燥合併之有機相，濃縮，得到8-溴咪唑并[1,2-b]嗒

-7-羧酸(1.25g，92.99%產率)，為黃色固體。 To 8-isopropylimidazo[1,2-b]

To a THF (20 mL) solution of -7-carboxylic acid ethyl ester 7a (1.5 g, 5.55 mmol), a water (10 mL) solution of lithium hydroxide monohydrate (699.2 mg, 16.66 mmol) was added. The mixture was stirred at room temperature for 5 hours. After removing the solvent, the residue was acidified with HCl (2M) to pH=3~4. The mixture was extracted with EtOAc and the combined organic phases were dried and concentrated to give 8-bromoimidazo[1,2-b]

-7-carboxylic acid (1.25 g, 92.99% yield), as a yellow solid.

LC-MS: m/z 241.1 [M+H] ⁺ .

Step 2

-7-羧酸17a(1.25g，5.16mmol)之甲苯(15mL)與2-甲基丙-2-醇(10mL)攪拌溶液中，添加三乙胺(2.61g，25.82mmol，、二苯基磷醯基疊氮化物(1.71g，6.20mmol，1.34mL)、三級丁氧羰酐(1.69g，7.75mmol，1.78mL)。於100℃攪拌反應混合物4小時。真空去除溶劑後，殘留物利用快速管柱層析法純化(以己烷類/乙酸乙酯=1：1洗提)，得到N-(8-溴咪唑并[1,2-b]嗒

-7-基)胺甲酸三級丁酯(820mg，41.43%產率)，為白色固體。 To 8-bromoimidazo[1,2-b]

To the stirred solution of -7-carboxylic acid 17a (1.25g, 5.16mmol) in toluene (15mL) and 2-methylpropan-2-ol (10mL), add triethylamine (2.61g, 25.82mmol, diphenyl Phosphoryl azide (1.71g, 6.20mmol, 1.34mL), tertiary butoxycarbonyl anhydride (1.69g, 7.75mmol, 1.78mL). The reaction mixture was stirred at 100°C for 4 hours. After the solvent was removed in vacuo, the residue Purify by flash column chromatography (eluting with hexanes/ethyl acetate=1:1) to obtain N-(8-bromoimidazo[1,2-b]

-7-yl) carbamic acid tertiary butyl ester (820 mg, 41.43% yield) as a white solid.

LC-MS: m/z 313.1 [M+H] ⁺ .

Step 3

-7-基)胺甲酸三級丁酯18a(800mg，2.55mmol)之MeOH(10mL)攪拌溶液中，添加三乙胺(1.29g，12.77mmol)、[1,1'-雙(二苯膦基)二茂鐵]二氯鈀(II)(186.9mg，0.255mmol)。CO(3MPa)下，於100℃攪拌所得反應混合物16小時。冷卻反應混合物至室溫。真空去除溶劑後，殘留物利用快速管柱層析法(以己烷類/乙酸乙酯=1：1洗提)純化，得到7-(三級丁氧羰胺基)咪唑并[1,2-b]嗒

-8-羧酸甲酯(460mg，61.60%產率)，為黃色固體。 To N-(8-bromoimidazo[1,2-b]

-7-yl) carbamic acid tertiary butyl 18a (800mg, 2.55mmol) in MeOH (10mL) stirred solution, add triethylamine (1.29g, 12.77mmol), [1,1'-bis(diphenylphosphine) Yl)ferrocene]dichloropalladium(II) (186.9 mg, 0.255 mmol). The resulting reaction mixture was stirred at 100°C for 16 hours under CO (3MPa). The reaction mixture was cooled to room temperature. After the solvent was removed in vacuo, the residue was purified by flash column chromatography (eluted with hexanes/ethyl acetate=1:1) to obtain 7-(tertiary butoxycarbonylamino)imidazo[1,2 -b] Da

Methyl-8-carboxylate (460 mg, 61.60% yield), as a yellow solid.

LC-MS: m/z 293.1 [M+H] ⁺ .

Step 4

-8-羧酸甲酯19a(460mg，1.57mmol)之MeOH(10mL)攪拌溶液中，分數次添加硼氫化鈉(297mg，7.87mmol)，於25℃攪拌所得反應混合物16小時。以飽和NH₄Cl猝滅反應並以EtOAc(10mL)萃取該混合物，合併之有機相以鹽水(5mL)洗滌，以無水Na₂SO₄乾燥， 過濾並真空濃縮。粗產物利用快速管柱層析法(DCM/MeOH=20：1)純化，得到N-[8-(羥甲基)咪唑并[1,2-b]嗒

-7-基]胺甲酸三級丁酯(300mg，72.13%產率)，為黃色固體。 At 0℃, at 7-(tertiary butoxycarbonylamino) imidazo[1,2-b]

To the stirring solution of methyl-8-carboxylate 19a (460 mg, 1.57 mmol) in MeOH (10 mL), sodium borohydride (297 mg, 7.87 mmol) was added several times, and the resulting reaction mixture was stirred at 25°C for 16 hours. The reaction was quenched with saturated NH ₄ Cl and the mixture was extracted with EtOAc (10 mL), the combined organic phase was washed with brine (5 mL), dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by flash column chromatography (DCM/MeOH=20:1) to obtain N-[8-(hydroxymethyl)imidazo[1,2-b]

Tertiary butyl -7-yl]carbamate (300 mg, 72.13% yield) as a yellow solid.

LC-MS: m/z 265.1 [M+H] ⁺ .

Step 5

-7-基]胺甲酸三級丁酯19b(300mg，1.14mmol)之MeOH(5.00mL)攪拌溶液中，添加亞硫醯氯(1.35g，11.35mmol)，然後於60℃攪拌所得反應混合物16小時。真空去除溶劑後，殘留物利用快速管柱層析法(以DCM/MeOH=10：1洗提)純化，得到7-胺基咪唑并[1,2-b]嗒

-8-羧酸甲酯(90mg，41.05%產率)，為黃色固體。 At 0℃, in N-[8-(hydroxymethyl)imidazo[1,2-b]

-7-yl] carbamic acid tertiary butyl ester 19b (300 mg, 1.14 mmol) in MeOH (5.00 mL) was added to a stirred solution of thiol chloride (1.35 g, 11.35 mmol), and then the resulting reaction mixture was stirred at 60°C 16 hour. After the solvent was removed in vacuo, the residue was purified by flash column chromatography (eluted with DCM/MeOH=10:1) to obtain 7-aminoimidazo[1,2-b]

Methyl-8-carboxylate (90 mg, 41.05% yield), as a yellow solid.

LC-MS: m/z 179.1 [M+H] ⁺ .

Step 6

-7-胺19b(90mg，0.505mmol)之DCM(2.00mL)溶液，於室溫攪拌反應混合物2小時。真空去除溶劑後，殘留物利用快速管柱層析法(以DCM：MeOH=30：1洗提)純化，得到1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[8-(甲氧基甲基)咪唑并[1,2-b]嗒

-7-基]脲(17.8mg，8.82%產率)，為白色固體。 To, 5-chloro-6- (triazol-2-yl) pyridin-3-amine 10a (90mg, 468.33 μ mol) of toluene (10 mL) was stirred solution was added triethylamine (153mg, 1.52mmol), bis carbonate (Trichloromethyl) ester (74.9 mg, 0.252 mmol). The reaction mixture was stirred at 100°C for 2 hours, and then at 0°C, 8-(methoxymethyl)imidazo[1,2-b] was added

A solution of -7-amine 19b (90 mg, 0.505 mmol) in DCM (2.00 mL), and the reaction mixture was stirred at room temperature for 2 hours. After the solvent was removed in vacuo, the residue was purified by flash column chromatography (eluted with DCM:MeOH=30:1) to obtain 1-[5-chloro-6-(triazol-2-yl)-3-pyridine Yl]-3-[8-(methoxymethyl)imidazo[1,2-b]ta

-7-yl]urea (17.8 mg, 8.82% yield) as a white solid.

LC-MS: m/z 400.1[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 10.30(s, 1H), 9.04(s, 1H), 8.77(s, 1H), 8.55( d, J =2.4Hz,1H), 8.50(d, J =2.4Hz,1H), 8.22(d, J =1.2Hz,1H), 8.16(s,2H), 7.69(d, J =1.2Hz, 1H), 4.94 (s, 2H), 3.39 (s, 3H).

-7-基)脲之合成 Example 54. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(cyclopropylamine) Base) imidazo[1,2-b]ta

-7-yl) urea synthesis

Example 54 was prepared in a similar procedure to that described in Preparation Example 21 by using cyclopropylamine instead of oxetane-3-amine as the starting material.

LC-MS: m/z 445.0[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 9.68(s,1H), 8.58(d, J =2.4Hz,1H), 8.44(d, J = 2.4Hz, 1H), 8.36 (s, 1H), 8.19 (s, 1H), 8.14 (s, 2H), 8.06 (s, 1H), 7.18 (s, 1H), 3.36-3.29 (m, 1H) ), 0.76-0.69 (m, 4H).

-8-基)乙醯胺之合成 Example 55. N-(7-(3-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)ureido)imidazo[1,2 -b] Da

-8-yl) acetamide synthesis

step 1

-7-羧酸乙酯7a(4g，14.81mmol)之THF(50mL)與水(30mL)溶液中，添加LiOH(1.24g，29.62mmol)。於室溫攪拌所得混合物10分鐘。TLC顯示反應完成。以1M HCl酸化該混合物至pH=6，以EtOAc萃取。乾燥合併之有機層，過濾及濃縮，得到8-溴咪唑并[1,2-b]嗒

-7-羧酸(3.26g，13.47mmol，91%產率)，為黃色固體。LC-MS：m/z 241.9[M+H]⁺。 To 8-bromoimidazo[1,2-b]

To a solution of ethyl-7-carboxylate 7a (4g, 14.81mmol) in THF (50mL) and water (30mL), LiOH (1.24g, 29.62mmol) was added. The resulting mixture was stirred at room temperature for 10 minutes. TLC showed that the reaction was complete. The mixture was acidified with 1M HCl to pH=6, and extracted with EtOAc. The combined organic layer was dried, filtered and concentrated to obtain 8-bromoimidazo[1,2- b ]

-7-carboxylic acid (3.26 g, 13.47 mmol, 91% yield) as a yellow solid. LC-MS: m/z 241.9 [M+H] ⁺ .

Step 2

-7-羧酸17a(3.26g，13.47mmol)、[疊氮(苯基)磷醯基]苯(3.93g，16.16mmol)、N,N-二乙基乙胺(6.81g，67.35mmol，9.39mL)、Boc₂O(14.70g，67.35mmol)於甲苯(25mL)與2-甲基丙-2-醇(25mL)中之混合物脫氣並再灌滿N₂(三次)。於100℃加熱混合物12小時後，LCMS顯示反應完成。濃縮該混合物，以EA(100mL)稀釋殘留物，以鹽水(40mLx2)洗滌，並乾燥。濃縮有機層，粗產物利用快速矽膠層析法(石油醚/乙酸乙酯=2/1)純化，得到N-(8-溴咪唑并[1,2-b]嗒

-7-基)胺甲酸三級丁酯(2.3g，55%產率)，為黃色固體。LC-MS：m/z 315.0[M+H]⁺。 Make 8-bromoimidazo[1,2- b ]

-7-carboxylic acid 17a (3.26g, 13.47mmol), [azido(phenyl)phosphinyl]benzene (3.93g, 16.16mmol), N , N -diethylethylamine (6.81g, 67.35mmol, 9.39 mL), Boc ₂ O (14.70 g, 67.35 mmol) in toluene (25 mL) and 2-methylpropan-2-ol (25 mL) were degassed and refilled with N ₂ (three times). After heating the mixture at 100°C for 12 hours, LCMS showed that the reaction was complete. The mixture was concentrated, and the residue was diluted with EA (100 mL), washed with brine (40 mL×2), and dried. The organic layer was concentrated, and the crude product was purified by flash silica gel chromatography (petroleum ether/ethyl acetate=2/1) to obtain N-(8-bromoimidazo[1,2-b]

Tertiary butyl -7-yl)carbamate (2.3 g, 55% yield) as a yellow solid. LC-MS: m/z 315.0 [M+H] ⁺ .

Step 3

-7-基)胺甲酸三級丁酯18a(850mg，2.71mmol)之N,N-二甲基甲醯胺(15mL)溶液中，添加Cs₂CO₃(1.77g，5.43mmol)。於80℃攪拌所得混合物2小時。TLC顯示反應完成。以EA(50mL)稀釋該混合物並予以過濾。濾液以飽和NH₄Cl(20mL)、1M LiCl(20mLx2)洗滌，並濃縮。殘留物利用矽膠管柱層析法純化(石油醚/乙酸乙酯=3/1)，得到N-(8-溴咪唑并[1,2-b]嗒

-7-基)-N-[(4-甲氧基苯基)甲基]胺甲酸三級丁酯(1.1g，94%產率)，為白色固體。 To N-(8-bromoimidazo[1,2-b]

-7-yl) carbamic acid tertiary butyl 18a (850 mg, 2.71 mmol) in N , N -dimethylformamide (15 mL) solution, add Cs ₂ CO ₃ (1.77 g, 5.43 mmol). The resulting mixture was stirred at 80°C for 2 hours. TLC showed that the reaction was complete. The mixture was diluted with EA (50 mL) and filtered. The filtrate was _{washed with saturated NH 4} Cl (20 mL), 1M LiCl (20 mL×2), and concentrated. The residue was purified by silica gel column chromatography (petroleum ether/ethyl acetate=3/1) to obtain N-(8-bromoimidazo[1,2-b]

-7-yl)-N-[(4-methoxyphenyl)methyl]carbamic acid tertiary butyl ester (1.1 g, 94% yield), a white solid.

LC-MS: m/z 433.1 [M+H] ⁺ .

Step 4

-7-基)-N-[(4-甲氧基苯基)甲基]胺甲酸三級丁酯18b(1.1g，2.54mmol)、乙醯胺(299mg，5.08mmol)、Cs₂CO₃(1.65g，5.08mmol)、Pd₂(dba)₃(465mg，507.73μmol)與黃磷(587mg，1.02mmol)於1,4-二

烷(20mL)中之混合物脫氣並以N₂回填(三次)。混合物於100℃加熱12小時後，LCMS顯示反應完成。過濾去除沉澱物並濃縮濾液。殘留物利用製備型TLC純化，得到 N-(8-乙醯胺基咪唑并[1,2-b]嗒

-7-基)-N-[(4-甲氧基苯基)甲基]胺甲酸三級丁酯(600mg，57%產率)，為黃色固體。LC-MS：m/z 412.2[M+H]⁺。 Make N-(8-bromoimidazo[1,2-b]

-7-yl)-N-[(4-methoxyphenyl)methyl]carbamic acid tertiary butyl ester 18b (1.1g, 2.54mmol), acetamide (299mg, 5.08mmol), Cs ₂ CO _{3 (1.65g, 5.08mmol), Pd 2} (dba) 3 (465mg, 507.73 μ mol) and phosphorus (587mg, 1.02mmol) in 1,4

The mixture in alkane (20 mL) was degassed and _{backfilled with N 2} (three times). After the mixture was heated at 100°C for 12 hours, LCMS showed that the reaction was complete. The precipitate was removed by filtration and the filtrate was concentrated. The residue was purified by preparative TLC to obtain N -(8-acetamidoimidazo[1,2-b]

-7-yl)-N-[(4-methoxyphenyl)methyl]carbamic acid tertiary butyl ester (600 mg, 57% yield), as a yellow solid. LC-MS: m/z 412.2 [M+H] ⁺ .

Step 5

-7-基)-N-[(4-甲氧基苯基)甲基]胺甲酸三級丁酯19c(600mg，1.46mmol)之TFA(5mL)溶液12小時。濃縮該混合物，殘留物以DCM(30mL)稀釋，以飽和NaHCO₃(15mLx2)洗滌，然後濃縮，得到粗產物，利用矽膠層析法(DCM/MeOH=20/1)將其純化，得到N-(7-胺基咪唑并[1,2-b]嗒

-8-基)乙醯胺(180mg，65%產率)，為黃色固體。LC-MS：m/z 192.1[M+H]⁺。 At room temperature, stir N -(8-acetamidoimidazo[1,2-b]

-7-yl)-N-[(4-methoxyphenyl)methyl]carbamic acid tertiary butyl ester 19c (600 mg, 1.46 mmol) in TFA (5 mL) for 12 hours. The mixture was concentrated, the residue was diluted with DCM (30 mL), _{washed with saturated NaHCO 3} (15 mLx2), and then concentrated to obtain the crude product, which was purified by silica gel chromatography (DCM/MeOH=20/1) to obtain N- (7-aminoimidazo[1,2- b ]

-8-yl)acetamide (180 mg, 65% yield), as a yellow solid. LC-MS: m/z 192.1 [M+H] ⁺ .

Step 6

-8-基)乙醯胺20b(50mg，0.261mmol)與N-[5-氯-6-(三唑-2-基)-3-吡啶基]胺甲酸苯酯(99mg，0.314mmol)之DMF溶液12小時。LCMS顯示反應完全。以EtOAc(15mL)稀釋該混合物，以1M LiCl洗滌並濃縮。殘留物利用製備型HPLC純化，得到N-[7-[[5-氯-6-(三唑-2-基)-3-吡啶基]胺甲醯胺基]咪唑并[1,2-b]嗒

-8-基]乙醯胺(23.8mg，22%產率)，為白色固體。LC-MS：m/z 413.1[M+H]⁺；¹H NMR(400MHz,DMSO-d ₆)：δ 10.90(s,1H),10.41(s,1H),8.83(s,1H),8.54(d,J=2.4Hz,1H),8.45(d,J=2.4Hz,1H),8.26(s,1H),8.15(s,3H),7.70(s,1H),2.30(s,3H)。 At 80℃, stir N -(7-aminoimidazo[1,2- b ]

-8-yl)acetamide 20b (50mg, 0.261mmol) and N- [5-chloro-6-(triazol-2-yl)-3-pyridyl]carbamate (99mg, 0.314mmol) DMF solution for 12 hours. LCMS showed that the reaction was complete. The mixture was diluted with EtOAc (15 mL), washed with 1M LiCl and concentrated. The residue was purified by preparative HPLC to obtain N -[7-[[5-chloro-6-(triazol-2-yl)-3-pyridyl]aminocarboxamido]imidazo[1,2- b ]despair

-8-yl]acetamide (23.8 mg, 22% yield) as a white solid. LC-MS: m/z 413.1[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.90 (s, 1H), 10.41 (s, 1H), 8.83 (s, 1H), 8.54 (d, J =2.4Hz,1H),8.45(d, J =2.4Hz,1H),8.26(s,1H),8.15(s,3H),7.70(s,1H),2.30(s,3H) .

-8-羧醯胺之合成 Example 56. 2-Chloro-7-(3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)ureido)-N,N- Dimethylimidazo[1,2-b]ta

Synthesis of -8-Carboxamide

step 1

-8-羧酸甲酯19d1(200mg，612.12μmol)之THF(4.0mL)攪拌溶液中，添加LiOH(1.0M，1.8mL)，攪拌2小時並經LCMS判斷反應完全，然後添加3M HCl以調至pH=2至3並以EA(40mL)萃取，其有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，得到7-(三級丁氧基羰胺基)-2-氯-咪唑并[1,2-b]嗒

-8-羧酸(190mg，607.59μmol，99.26%產率)，LC-MS：m/z 313[M+H]⁺，粗產物直接用於下一步驟不需進一步純化。 At 7-(tertiary butoxycarbonylamino)-2-chloro-imidazo[1,2-b]ta

8-carboxylate 19d1 (200mg, 612.12 μ mol) of THF (4.0mL) was stirred solution was added LiOH (1.0M, 1.8mL), stirred for 2 h and judged complete by LCMS, 3M HCl was then added to Adjusted to pH=2 to 3 and extracted with EA (40 mL), the organic phase was washed with brine (5 mL), _{dried with Na 2} SO ₄ , filtered and concentrated in vacuo to obtain 7-(tertiary butoxycarbonylamino) -2-Chloro-imidazo[1,2-b]

8-carboxylic acid (190mg, 607.59 μ mol, 99.26 % yield), LC-MS: m / z 313 [M + H] +, the crude product was directly used in the next step without further purification.

Step 2

-8-羧酸19d2(190mg，607.59μmol)之DMF(2.0mL)攪拌溶液中，添加二甲胺鹽酸鹽(99.1mg，1.22mmol)、HATU(462mg，1.22mmol)、DIEA(471mg，3.65mmol)，攪拌4.0小時，經LCMS判斷反應完全，添加4.0mL水，以EA(40mL)萃取，有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，粗混合物利用矽膠管柱層析法純化，使用EA/PE=0：100-1：10作為洗提液，得到N-[2-氯-8-(二甲基胺甲醯基)咪唑并[1,2-b]嗒

-7-基]胺甲酸三級丁酯(150mg，441.47μmol，72.66%產率)，LC-MS：m/z 340[M+H]⁺。 At 7-(tertiary butoxycarbonylamino)-2-chloro-imidazo[1,2-b]ta

8-carboxylic acid 19d2 (190mg, 607.59 μ mol) of DMF (2.0mL) was stirred solution was added dimethylamine hydrochloride (99.1mg, 1.22mmol), HATU ( 462mg, 1.22mmol), DIEA (471mg, 3.65mmol), stirred for 4.0 hours, the reaction was judged complete by LCMS, 4.0mL of water was added, extracted with EA (40mL), the organic phase was washed with brine (5mL), _{dried with Na 2} SO ₄ , filtered and concentrated in vacuo. The crude mixture was used Purified by silica gel column chromatography, using EA/PE=0:100-1:10 as the eluent to obtain N-[2-chloro-8-(dimethylaminomethanoyl)imidazo[1,2 -b] Da

7-yl] butyl amine three acid (150mg, 441.47 μ mol, 72.66 % yield), LC-MS: m / z 340 [M + H] +.

Step 3

-7-基]胺甲酸三級丁酯19d3(150mg，441.47μmol)攪拌溶液中，添加HCl/EA(1.0M，5mL)，所得混合物攪拌隔夜，以Na₂CO₃調到pH=9至10並以EA(40mL)萃取，其有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，得到7-胺基-2-氯-N,N-二甲基-咪唑并[1,2-b]嗒

-8-羧醯胺(100mg，417.26μmol，94.52%產率)，LC-MS：m/z 240[M+H]⁺，粗產物直接用於下一步驟不需進一步純化。 To N-[2-Chloro-8-(dimethylaminocarboxyl)imidazo[1,2-b]ta

-7-yl] carbamic acid tertiary butyl ester 19d3 (150 mg, 441.47 μ mol) was added to the stirring solution, HCl/EA (1.0 M, 5 mL) was added, the resulting mixture was stirred overnight, and the pH was adjusted to 9 _{with Na 2} CO ₃ 10 and extracted with EA (40 mL), the organic phase was washed with brine (5 mL), _{dried over Na 2} SO ₄ , filtered and concentrated in vacuo to obtain 7-amino-2-chloro-N,N-dimethyl-imidazole And [1,2-b]

-8- 2carboxamide (100mg, 417.26 μ mol, 94.52 % yield), LC-MS: m / z 240 [M + H] +, the crude product was directly used in the next step without further purification.

Step 4

-8-羧醯胺20c(50mg，208.63μmol)之DMF(4.0mL)攪拌溶液中，添加CDI(120mg，834.51μmol)與TEA(126.7mg，1.25mmol，174.47uL)，於80℃加熱反應混合物4小時，冷卻至40℃。於其內添加5-氯-6-(三唑-2-基)吡啶-3-胺(40.8mg，208.63μmol)。此混合物於80℃加熱隔夜，冷卻至室溫並以EA(40mL)稀釋，其有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，粗混合物利用製備型HPLC純化，得到2-氯-7-[[5-氯-6-(三唑-2-基)-3-吡啶基]胺甲醯胺基]-N,N-二甲基-咪唑并[1,2-b]嗒

-8-羧醯胺(6.0mg，13.01μmol，6.23%產率)。LC-MS：m/z 462[M+H]⁺；¹H NMR(400MHz,DMSO-d6)δ 9.16(s,1H),8.55(d,J=2.3Hz,1H),8.47(d,J=2.3Hz,1H),8.44(s,1H),8.16(s,2H),3.12(s,3H),2.89(s,3H)。 To 7-amino-2-chloro-N,N-dimethyl-imidazo[1,2-b]

-8- 2carboxamide 20c (50mg, 208.63 μ mol) of DMF (4.0mL) was stirred solution was added CDI (120mg, 834.51 μ mol) and TEA (126.7mg, 1.25mmol, 174.47uL) , heated at 80 deg.] C The reaction mixture was cooled to 40°C for 4 hours. Therein of 5-chloro-6- (triazol-2-yl) pyridin-3-amine (40.8mg, 208.63 μ mol). The mixture was heated at 80°C overnight, cooled to room temperature and diluted with EA (40 mL). The organic phase was washed with brine (5 mL), _{dried over Na 2} SO ₄ , filtered and concentrated in vacuo. The crude mixture was purified by preparative HPLC. Obtain 2-chloro-7-[[5-chloro-6-(triazol-2-yl)-3-pyridyl]aminocarboxamido]-N,N-dimethyl-imidazo[1,2 -b] Da

-8-Carboxamide (6.0 mg, 13.01 μ mol, 6.23% yield). LC-MS: m/z 462[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d6 ) δ 9.16(s,1H), 8.55(d, J =2.3Hz,1H), 8.47(d, J =2.3Hz, 1H), 8.44 (s, 1H), 8.16 (s, 2H), 3.12 (s, 3H), 2.89 (s, 3H).

-8-羧酸之合成 Example 57. 2-Chloro-7-(3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)ureido)imidazo[1, 2-b]

Synthesis of -8-carboxylic acid

step 1

-7-羧酸乙酯7c(300mg，985.13μmol)、草酸二乙酯(172.8mg,1.18mmol，159.96uL)、N,N-二甲基吡啶-4-胺(144.4mg，1.18mmol)與EtOH(6.0mL)，N₂下，攪拌反應混合物10分鐘，添加氯化雙(三苯膦)鈀(II)(69.2mg，98.51μmol)，微波輻射下，於120℃加熱0.5小時。令混合物冷卻至室溫。過濾並蒸發溶劑後，粗混合物利用矽膠管柱層析法純化，使用EA/PE=0：100-100：0作為洗提液，得到2-氯-7-乙氧羥基-咪唑并[1,2-b]嗒

-8-羧酸(190mg，704.64μmol，71.53%產率)，LC-MS：m/z 270[M+H]⁺。 Add 8-bromo-2-chloro-imidazo[1,2-b]ta to the microwave bottle

7-carboxylic acid ethyl ester 7c (300mg, 985.13 μ mol) , diethyl oxalate (172.8mg, 1.18mmol, 159.96uL), N, N- dimethyl-pyridin-4-amine (144.4mg, 1.18mmol) and EtOH (6.0mL), under N _2, the reaction mixture was stirred for 10 minutes, bis (triphenylphosphine) palladium (II) (69.2mg, 98.51 μ mol), under microwave irradiation, heating at 120 ℃ 0.5 h. The mixture was allowed to cool to room temperature. After filtering and evaporating the solvent, the crude mixture was purified by silica gel column chromatography, using EA/PE=0:100-100:0 as the eluent to obtain 2-chloro-7-ethoxyhydroxy-imidazo[1, 2-b]

8-carboxylic acid (190mg, 704.64 μ mol, 71.53 % yield), LC-MS: m / z 270 [M + H] +.

Step 2

-8-羧酸8q(100mg，370.86μmol)之THF(2.0mL)攪拌溶液中，添加LiOH(1.0M，1.0ml)，攪拌混合物5小時，經LCMS判斷反應完全，然後添加3M鹽酸調至pH=2至3並以EA(40mL)萃取，有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，得到2-氯咪唑并[1,2-b]嗒

-7,8-二羧酸(85mg，351.84μmol，94.87%產率)，LC-MS：m/z 242[M+H]⁺，粗產物直接用於下一步驟不需進一步純化。 On 2-chloro-7-ethoxyhydroxy-imidazo[1,2-b]ta

8-carboxylic acid 8q (100mg, 370.86 μ mol) of THF (2.0mL) was stirred solution was added LiOH (1.0M, 1.0ml), the mixture was stirred for 5 hours, the reaction was judged complete by LCMS, and then 3M hydrochloric acid was adjusted to pH=2 to 3 and extracted with EA (40 mL), the organic phase was washed with brine (5 mL), _{dried with Na 2} SO ₄ , filtered and concentrated in vacuo to obtain 2-chloroimidazo[1,2-b]

7,8-dicarboxylic acid (85mg, 351.84 μ mol, 94.87 % yield), LC-MS: m / z 242 [M + H] +, the crude product was directly used in the next step without further purification.

Step 3

-7,8-二羧酸9q(85mg，351.84μmol)之1,4-二

烷(2.0mL)攪拌溶液中，添加DPPA(128.3mg，527.76μmol)與TEA(106.8mg，1.06mmol，147.12uL)。於室溫攪拌反應混合物2小時。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(82.6mg，422.21μmol)，於100℃加熱4小時，經LCMS判斷反應完全，添加1.0mL冰水，以EA(40mL)萃取，有機相以鹽水(5mL)洗滌，以Na₂SO₄乾燥，過濾並真空濃縮，粗混合物利用製備型HPLC純化，得到2-氯-7-[[5-氯-6-(三唑-2-基)-3-吡啶基]胺甲醯胺基]咪唑并[1,2-b]嗒

-8-羧酸(9.0mg，20.73μmol，5.89%產率)，為白色固體，LC-MS：m/z 435[M+H]⁺；¹H NMR(400MHz,DMSO-d6)δ 9.11(d,J=8.6Hz,1H),8.47(d,J=2.3Hz,1H),8.42(d,J=2.3Hz,1H),8.31(d,J=10.2Hz,1H),8.09(s,2H)。 At 2-chloroimidazo[1,2-b]

-7,8-Dicarboxylic acid 9q (85mg, 351.84 μ mol) of 1,4-bis

Dioxane (2.0 mL) was stirred solution was added DPPA (128.3mg, 527.76 μ mol) and TEA (106.8mg, 1.06mmol, 147.12uL) . The reaction mixture was stirred at room temperature for 2 hours. Then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (82.6 mg, 422.21 μ mol) was added and heated at 100°C for 4 hours. The reaction was judged to be complete by LCMS, and 1.0 mL ice water was added to EA (40 mL) was extracted, the organic phase was washed with brine (5 mL), _{dried over Na 2} SO ₄ , filtered and concentrated in vacuo. The crude mixture was purified by preparative HPLC to give 2-chloro-7-[[5-chloro-6- (Triazol-2-yl)-3-pyridyl]carboxamido]imidazo[1,2-b]ta

-8-carboxylic acid (9.0 mg, 20.73 μ mol, 5.89% yield), white solid, LC-MS: m/z 435[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ) δ 9.11 (d, J =8.6Hz,1H),8.47(d, J =2.3Hz,1H),8.42(d, J =2.3Hz,1H),8.31(d, J =10.2Hz,1H),8.09(s ,2H).

-7-基)脲之合成 Example 58. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(imidazo[1,2-b]

-7-yl) urea synthesis

-7-羧酸酯作為起始物質，以類似製備實施例10所述程序製備實施例58。 By using 8-bromoimidazo[1,2- b ]

The -7-carboxylate was used as the starting material, and Example 58 was prepared in a similar procedure as described in Preparation Example 10.

LC-MS: m/z 356.0; ¹ H NMR (400MHz, DMSO- d ₆ ): δ 9.79 (s, 1H), 8.59 (s, 1H), 8.62-8.59 (m, 2H), 8.47-8.46 (m , 1H), 8.17-8.16 (m, 4H), 7.66 (s, 1H).

-7-基)-3-(5-氯-6-甲氧吡啶-3-基)脲之合成 Example 59. 1-(8-Bromo-2-chloroimidazo[1,2-b]

Synthesis of -7-yl)-3-(5-chloro-6-methoxypyridin-3-yl)urea

-7-羧酸酯作為起始物質，以類似製備實施例10所述程序製備實施例59。 By using 8-bromoimidazo[1,2- b ]

-7-carboxylate was used as the starting material, and Example 59 was prepared in a similar procedure to that described in Preparation Example 10.

LC-MS: m/z 430.8 [M+H] ⁺ .

-7-基)-3-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)脲之合成 Example 60. 1-(8-Amino-2-chloroimidazo[1,2-b]

Synthesis of -7-yl)-3-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)urea

-7-羧酸酯作為起始物質，以類似製備實施例10所述程序製備實施例60。 By using 8-bromoimidazo[1,2- b ]

-7-carboxylate was used as the starting material, and Example 60 was prepared in a similar procedure to that described in Preparation Example 10.

LC-MS: m/z 405[M+H] ⁺ ; ¹ H NMR(400MHz,DMSO- d ₆ ) δ 10.20(s,1H),8.85(s,1H),8.60(d, J =2.2Hz, 1H), 8.45 (d, J = 2.2 Hz, 1H), 8.19 (s, 1H), 8.17 (s, 1H), 8.14 (s, 2H), 6.98 (s, 2H).

-7-基]脲/(S)-1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲/(R)-1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲之合成 Examples 61 to 63. 1-[5-Chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl)imidazo[1,2 -b] Da

-7-yl]urea/( S )-1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl)imidazole And [1,2-b]

-7-yl]urea/( R )-1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl)imidazole And [1,2-b]

Synthesis of -7-yl]urea

step 1

-7-基)胺甲酸三級丁酯18a(10.36g，33.08mmol)之N,N-二甲基甲醯胺(50mL)溶液中，添加三丁基(1-乙氧基乙烯基)錫烷(23.90g，66.17mmol，22.33mL)，隨後於N₂下，添加氯化雙(三苯膦)鈀(II)(2.32g，3.31mmol)。所得反應混合物於100℃攪拌3小時。反應完全後，濃縮混合物，殘留物利用快速管柱層析法(石油醚：乙酸乙酯3：1)純化，得到N-[8-(1-乙氧基乙烯基)咪唑并[1,2-b]嗒

-7-基]胺甲酸三級丁酯(10.06g，99%產率)，為黃色油。 At room temperature, at N-(8-bromoimidazo[1,2-b]

-7-yl) carbamic acid tertiary butyl 18a (10.36g, 33.08mmol) in N,N-dimethylformamide (50mL) solution, add tributyl(1-ethoxyvinyl)tin Alkane (23.90 g, 66.17 mmol, 22.33 mL), then under N ₂ bis(triphenylphosphine)palladium(II) chloride (2.32 g, 3.31 mmol) was added. The resulting reaction mixture was stirred at 100°C for 3 hours. After the completion of the reaction, the mixture was concentrated, and the residue was purified by flash column chromatography (petroleum ether: ethyl acetate 3:1) to obtain N-[8-(1-ethoxyvinyl)imidazo[1,2 -b] Da

-7-yl] carbamic acid tertiary butyl ester (10.06 g, 99% yield) as a yellow oil.

LC-MS: m/z 305 [M+H] ⁺ .

Step 2

-7-基]胺甲酸三級丁酯19e(2g，6.57mmol)之二

烷(20mL)溶液中，添加HCl/二

烷(20mL)。於80℃攪拌混合物2小時，然後真空去除溶劑。接著添加HCl/水(20mL)，於80℃攪拌混合物2小時。然後以Na₂CO₃(aq)鹼化混合物至pH~8，過濾固體，乾燥，得到化合物1-(7-胺基咪唑并[1,2-b]嗒

-8-基)乙酮(1.15g，99.33%產率)，為黃色固體。 To N-[8-(1-ethoxyvinyl)imidazo[1,2-b]ta

-7-yl] carbamic acid tertiary butyl 19e (2g, 6.57mmol) bis

Alkane (20mL) solution, add HCl/two

Alkane (20 mL). The mixture was stirred at 80°C for 2 hours, and then the solvent was removed in vacuo. Then HCl/water (20 mL) was added, and the mixture was stirred at 80°C for 2 hours. Then basify the mixture with Na ₂ CO ₃ (aq) to pH~8, filter the solid, and dry to obtain the compound 1-(7-aminoimidazo[1,2-b]

-8-yl)ethanone (1.15 g, 99.33% yield) as a yellow solid.

LC-MS: m/z 177[M+H] ⁺ .

Step 3

-8-基)乙酮20d(4.8g，27.25mmol)之甲醇(60mL)溶液中，添加硼氫化鈉(2.06g，54.49mmol，1.93mL)。於室溫攪拌混合物4小時。反應完全後，以水(50mL)稀釋反應混合物並以EtOAc(3×250mL)萃取。合併之有機相以無水Na₂SO₄乾燥，過濾並真空濃縮。粗產物利用快速層析法(DCM/MeOH 15：1)純化，得到1-(7-胺基咪唑并[1,2-b]嗒

-8-基)乙醇(3.84g，79%產率)，為灰色固體。 At 0℃, at 1-(7-aminoimidazo[1,2-b]

To a solution of -8-yl) ethyl ketone 20d (4.8 g, 27.25 mmol) in methanol (60 mL), sodium borohydride (2.06 g, 54.49 mmol, 1.93 mL) was added. The mixture was stirred at room temperature for 4 hours. After the reaction was complete, the reaction mixture was diluted with water (50 mL) and extracted with EtOAc (3×250 mL). The combined organic phase was dried over anhydrous Na ₂ SO ₄ , filtered and concentrated in vacuo. The crude product was purified by flash chromatography (DCM/MeOH 15:1) to obtain 1-(7-aminoimidazo[1,2-b]

-8-yl) ethanol (3.84 g, 79% yield) as a gray solid.

LC-MS: m/z 179 [M+H] ⁺ .

Step 4

-8-基)乙醇21a(4g，22.45mmol)之甲醇(45.00mL)溶液中，添加亞硫醯氯(2.4mL)，於50℃攪拌混合物隔夜。反應完全後，濃縮反應混合物，利用快速層析法(DCM/MeOH 30：1)純化，得到8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-胺(3.6g，83%產率)，為黃色固體。 At room temperature, at 1-(7-aminoimidazo[1,2-b]

To a methanol (45.00 mL) solution of -8-yl)ethanol 21a (4 g, 22.45 mmol) was added sulfite chloride (2.4 mL), and the mixture was stirred at 50°C overnight. After the reaction is complete, the reaction mixture is concentrated and purified by flash chromatography (DCM/MeOH 30:1) to obtain 8-(1-methoxyethyl)imidazo[1,2-b]

-7-amine (3.6 g, 83% yield), as a yellow solid.

LC-MS: m/z 193 [M+H] ⁺ .

Step 5

-7-胺22a(10g，52.02mmol)之THF(200mL)溶液中添加N-[5-氯-6-(三唑-2-基)-3-吡啶基]胺甲酸苯酯(17.25g，54.63mmol)。此混合物於80℃攪拌3小時，LCMS顯示用盡大部分起始原料。去除溶劑，粗產物利用快速層析法(DCM/MeOH 30：1)純化，得到1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲(61)(15g，69.67%產率)，為白色固體。 At room temperature, in 8-(1-methoxyethyl)imidazo[1,2-b]

To a solution of -7-amine 22a (10g, 52.02mmol) in THF (200mL) was added N-[5-chloro-6-(triazol-2-yl)-3-pyridyl] phenyl carbamate (17.25g, 54.63mmol). The mixture was stirred at 80°C for 3 hours, LCMS showed that most of the starting material was used up. The solvent was removed, and the crude product was purified by flash chromatography (DCM/MeOH 30:1) to obtain 1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8- (1-Methoxyethyl) imidazo[1,2-b]

-7-yl]urea ( 61 ) (15 g, 69.67% yield) as a white solid.

Using Prep-Chiral-SFC with the following conditions (Daicel Chiralpak, IG 4.6mm*250mm*5um, 5 to 50% Hex in EtOH, 1.0 mL/min), chiral separation of compounds ( 61 ). The parts are evaporated to dryness to obtain the desired compound.

-7-基]脲(62)或(R)-1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲(63)。 Peak 1: ( S )-1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl)imidazo[1, 2-b]

-7-yl]urea (62 ) or ( R )-1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl) Base) imidazo[1,2-b]ta

-7-yl]urea ( 63 ).

(0.5，CH₃OH)； Residence time: 6.146 minutes; ee (%): >99%,

(0.5, CH ₃ OH);

-7-基]脲(63)或(S)-1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲(62)。 Peak 2: ( R )-1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl)imidazo[1, 2-b]

-7-yl]urea ( 63 ) or ( S )-1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-(1-methoxyethyl) Base) imidazo[1,2-b]ta

-7-yl]urea ( 62 ).

(0.5，CH₃OH)；LC-MS：m/z 414[M+H]⁺；¹H NMR(400MHz,DMSO-d6)：δ 10.60(s,1H),9.12(s,1H),8.81(s,1H),8.55(d,J=2.4Hz,1H),8.50(d,J=2.4Hz,1H),8.20(d,J=1.6Hz,1H),8.16(s,2H),7.67(d,J=1.2Hz,1H),5.33(q,J=6.8Hz,1H),3.34(s,3H),1.56(d,J=6.4Hz,3H)。 Residence time: 7.681 minutes, ee : >99%;

(0.5, CH ₃ OH); LC-MS: m/z 414[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 10.60(s, 1H), 9.12(s, 1H), 8.81 (s,1H),8.55(d, J =2.4Hz,1H),8.50(d, J =2.4Hz,1H),8.20(d, J =1.6Hz,1H),8.16(s,2H),7.67 (d, J =1.2Hz,1H), 5.33(q, J =6.8Hz,1H), 3.34(s,3H), 1.56(d, J =6.4Hz,3H).

-7-基)脲/(S)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲/(R)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲之合成 Examples 64 to 66: 1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(1- Methoxyethyl) imidazo[1,2-b]

-7-yl)urea/( S )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro- 8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl)urea/( R )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro- 8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl) urea synthesis

step 1:

-7-羧酸乙酯7c(31g，102.3mmol)之THF(500mL)溶液中，添加氫氧化鋰一水合物(8.6g，204.6mmol)之水200mL)溶液。於室溫攪拌所得混合物10分鐘。反應混合物以HCl(1N)調至pH=3~4予以猝滅，將其以EtOAc萃取，乾燥合併之有機相，濃縮，得到8-溴-2-氯咪唑并[1,2-b]嗒

-7-羧酸(28g，99.6%)，為黃色固體。 On 8-bromo-2-chloroimidazo[1,2-b]

To the THF (500 mL) solution of ethyl-7-carboxylate 7c (31 g, 102.3 mmol), a solution of lithium hydroxide monohydrate (8.6 g, 204.6 mmol) in water 200 mL) was added. The resulting mixture was stirred at room temperature for 10 minutes. The reaction mixture was quenched with HCl (1N) adjusted to pH=3~4, extracted with EtOAc, and the combined organic phases were dried and concentrated to give 8-bromo-2-chloroimidazo[1,2-b]

-7-carboxylic acid (28 g, 99.6%), as a yellow solid.

LC-MS: m/z 276 [M+H] ⁺ .

Step 2:

-7-羧酸17c(28g，101.8mmol)之三級丁醇(280mL)與甲苯(300mL)溶液中，添加二苯基磷醯基疊氮化物(39.2g，142.5mmol)、三乙胺(51.5g，509mmol)與二碳酸二(三級丁酯)(66.58g，305.4mmol)。於100℃攪拌反應混合物4小時。LC-MS顯示耗盡起始原料並檢測到所需質量。濃縮後，所得殘留物利用快速管柱層析法(以於石油中0%至10%之乙酸乙酯洗提)純 化，得到(8-溴-2-氯咪唑并[1,2-b]嗒

-7-基)胺甲酸三級丁酯(19.4g，產率：55.1%)，為黃色固體。 On 8-bromo-2-chloroimidazo[1,2-b]

-7-carboxylic acid 17c (28g, 101.8mmol) in tertiary butanol (280mL) and toluene (300mL) solution, add diphenylphosphoryl azide (39.2g, 142.5mmol), triethylamine ( 51.5 g, 509 mmol) and di(tertiary butyl) dicarbonate (66.58 g, 305.4 mmol). The reaction mixture was stirred at 100°C for 4 hours. LC-MS showed that the starting material was consumed and the required mass was detected. After concentration, the residue obtained was purified by flash column chromatography (eluted with 0% to 10% ethyl acetate in petroleum) to obtain (8-bromo-2-chloroimidazo[1,2-b] despair

Tertiary butyl -7-yl)carbamate (19.4 g, yield: 55.1%), as a yellow solid.

LC-MS: m/z 347 [M+H] ⁺ .

Step 3:

-7-基)胺甲酸三級丁酯18c(7.75g，22.4mmol)之N,N-二甲基甲醯胺(100mL)溶液中，添加三丁基(1-乙氧乙烯基)錫烷(16.22g，44.8mmol)，將所得混合物脫氣並與N₂交換兩次，添加氯化雙(三苯膦)鈀(II)(3.14g，4.48mmol)。於100℃攪拌反應混合物4小時。LC-MS顯示耗盡起始原料並檢測到所需質量。濃縮後，所得殘留物利用矽膠管柱層析法，以於石油醚中之0%至10%乙酸乙酯洗提予以純化，得到(2-氯-8-(1-乙氧基乙烯基)咪唑并[1,2-b]嗒

-7-基)胺甲酸三級丁酯(5.9g，產率：77.9%)，為白色固體。 At room temperature, in (8-bromo-2-chloroimidazo[1,2-b]

-7-yl) carbamate tertiary butyl 18c (7.75g, 22.4mmol) in N,N-dimethylformamide (100mL) solution, add tributyl(1-ethoxyvinyl)stannane (16.22g, 44.8mmol), and the resulting mixture was degassed with N ₂ twice exchange, bis (triphenylphosphine) palladium (II) (3.14g, 4.48mmol) . The reaction mixture was stirred at 100°C for 4 hours. LC-MS showed that the starting material was consumed and the required mass was detected. After concentration, the residue obtained was purified by silica gel column chromatography and eluted with 0% to 10% ethyl acetate in petroleum ether to obtain (2-chloro-8-(1-ethoxyvinyl) Imidazo[1,2-b]

-7-yl) carbamic acid tertiary butyl ester (5.9 g, yield: 77.9%) as a white solid.

LC-MS: m/z 339 [M+H] ⁺ .

Step 4:

-7-基)胺甲酸三級丁酯19f(5.9g，17.46mmol)之1,4-二

烷(100mL)溶液中，添加HCl/1,4-二

烷(30mL)。於80℃攪拌反應混合物2小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮反應混合物，殘留物以三乙胺鹼化至pH~7至8。濃縮後，殘留物利用矽膠管柱層析法，以於石油醚中0%至30%之乙酸乙酯洗提予以純化，得到1-(7-胺基-2-氯咪唑并[1,2-b]嗒

-8-基)乙酮(2.0g，產率：54.5%)，為黃色固體。 On (2-chloro-8-(1-ethoxyvinyl)imidazo[1,2-b]ta

-7-yl) tertiary butyl carbamate 19f (5.9g, 17.46mmol) of 1,4-bis

In the alkane (100mL) solution, add HCl/1,4-bis

Alkane (30 mL). The reaction mixture was stirred at 80°C for 2 hours. The starting material was consumed and the required quality was detected by LC-MS. The reaction mixture was concentrated, and the residue was basified to pH~7-8 with triethylamine. After concentration, the residue was purified by silica gel column chromatography and eluted with 0% to 30% ethyl acetate in petroleum ether to obtain 1-(7-amino-2-chloroimidazo[1,2 -b] Da

-8-yl)ethanone (2.0 g, yield: 54.5%), as a yellow solid.

LC-MS: m/z 211 [M+H] ⁺ .

Step 5:

-8-基)乙酮20e(2.0g，9.52mmol)之甲醇(50mL)溶液中，添加NaBH₄(1.08g.28.57mmol)。於室溫攪拌所得混合物4小時。反應完全後，濃縮混合物，殘留物利用快速層析法，以甲醇/二氯甲烷=10/1洗提予以純化，得到1-(7-胺基-2-氯咪唑并[1,2-b]嗒

-8-基)乙醇(1.94g，產率：96%)，為黃色固體。 0℃, at 1-(7-amino-2-chloroimidazo[1,2-b]

To a solution of -8-yl) ethyl ketone 20e (2.0 g, 9.52 mmol) in methanol (50 mL), NaBH ₄ (1.08 g. 28.57 mmol) was added. The resulting mixture was stirred at room temperature for 4 hours. After the reaction is complete, the mixture is concentrated, and the residue is purified by flash chromatography with methanol/dichloromethane=10/1 to obtain 1-(7-amino-2-chloroimidazo[1,2-b ]despair

-8-yl) ethanol (1.94 g, yield: 96%) as a yellow solid.

LC-MS: m/z 213 [M+H] ⁺ .

Step 6:

-8-基)乙醇21c(1.94g，9.151mmol)之甲醇(60mL溶液中，添加SOCl₂(12mL)。所得混合物於50℃攪拌隔夜。耗盡起始原料並以LC-MS檢測到所需質量。濃縮所得混合物，殘留物利用矽膠管柱層析法，以於石油醚中0%至30%之乙酸乙酯洗提予以純化，得到2-氯-8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-胺(1.7g，產率：82.1%)，為黃色固體。 0℃, at 1-(7-amino-2-chloroimidazo[1,2-b]

-8-yl)ethanol 21c (1.94g, 9.151mmol) in methanol (60mL solution, add SOCl ₂ (12mL). The resulting mixture was stirred at 50°C overnight. The starting material was consumed and the required was detected by LC-MS Quality. The resulting mixture is concentrated, and the residue is purified by silica gel column chromatography and eluted with 0% to 30% ethyl acetate in petroleum ether to obtain 2-chloro-8-(1-methoxyethyl) )Imidazo[1,2-b]ta

-7-amine (1.7 g, yield: 82.1%), as a yellow solid.

LC-MS: m/z 227 [M+H] ⁺ .

Step 7:

-7-胺22c(300mg，1.327mmol)之N,N-二甲基甲醯胺(14mL)溶液中，添加(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)胺甲酸苯酯23a(752.6mg，2.389mmol)。微波輻射下，於90℃加熱所得混合物8小時。耗盡起始原料並以LC-MS檢測到所需質量。濃縮混合物，殘留物利用快速層析法(以甲醇/二氯甲烷：20/1洗提)純化，得到1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲(64)(401mg，產率：67.6%)，為白色固體。 To 2-chloro-8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-amine 22c (300mg, 1.327mmol) in N,N-dimethylformamide (14mL) solution, add (5-chloro-6-(2H-1,2,3-triazole-2- (Yl)pyridin-3-yl)phenylcarbamate 23a (752.6 mg, 2.389 mmol). Under microwave irradiation, the resulting mixture was heated at 90°C for 8 hours. The starting material was consumed and the required quality was detected by LC-MS. The mixture was concentrated, and the residue was purified by flash chromatography (eluted with methanol/dichloromethane: 20/1) to obtain 1-(5-chloro-6-(2H-1,2,3-triazole-2- Yl)pyridin-3-yl)-3-(2-chloro-8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 64 ) (401 mg, yield: 67.6%), as a white solid.

Using Prep-Chiral-SFC with the following conditions (Daicel Chiralpak, IG 4.6mm*250mm*5um, 5 to 50% Hex in EtOH, 1.0 mL/min), chiral separation of compounds ( 64 ). The parts are evaporated to dryness to obtain the desired compound.

-7-基)脲(65)或(R)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲(66)。 Peak 1: ( S )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(1 -Methoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 65 ) or ( R )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2 -Chloro-8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 66 ).

Residence time: 10.280 minutes, ee (%): >99%.

-7-基)脲(66)或(S)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(2-氯-8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲(65)。 Peak 2: ( R )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(1 -Methoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 66 ) or ( S )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2 -Chloro-8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 65 ).

Retention time: 11.856 minutes, ee (%): >99%.

LC-MS: m/z 448 [M+H] ⁺ . LC-MS: m/z 448[M+H] ⁺ ; ¹ H NMR (400MHz; DMSO- d6 ): δ 10.62 (s, 1 H), 9.21 (s, 1 H), 8.85 (s, 1 H) ,8.55(d, J =2.4Hz,1 H),8.49(d, J =2.0Hz,1 H),8.39(s,1 H),8.15(s,2 H),5.19-5.24(m,1 H), 3.34 (s, 3 H), 1.55 (d, J = 6.8 Hz, 3 H).

-7-基)脲之合成 Example 67. 1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(2-chloro-8-(1-(two (Methylamino) ethyl) imidazo[1,2-b]

-7-yl) urea synthesis

step 1

-7-羧酸乙酯8r(50mg，0.187mmol)之甲醇(3mL)溶液中，添加N-甲基甲胺(25.3mg，0.56mmol)與氰基硼氫化鈉(23.5 mg，0.374mmol)。於室溫攪拌反應混合物隔夜。LC-MS顯示耗盡起始原料並檢測到所需質量。濃縮反應混合物，所得殘留物利用矽膠管柱層析法，以於石油醚中之0%至100%乙酸乙酯洗提予以純化，得到2-氯-8-[1-(二甲胺基)乙基]咪唑并[1,2-b]嗒

-7-羧酸(36mg，71.72%產率)。 To 8-Acetyl-2-chloro-imidazo[1,2-b]

To a solution of ethyl-7-carboxylate 8r (50 mg, 0.187 mmol) in methanol (3 mL), N -methylmethylamine (25.3 mg, 0.56 mmol) and sodium cyanoborohydride (23.5 mg, 0.374 mmol) were added. The reaction mixture was stirred at room temperature overnight. LC-MS showed that the starting material was consumed and the required mass was detected. The reaction mixture was concentrated, and the residue obtained was purified by silica gel column chromatography and eluted with 0% to 100% ethyl acetate in petroleum ether to obtain 2-chloro-8-[1-(dimethylamino) Ethyl]imidazo[1,2- b ]ta

-7-carboxylic acid (36 mg, 71.72% yield).

LC-MS: m/z 269 [M+H] ⁺ .

Step 2

-7-羧酸9r(36mg，0.134mmol)之1,4-二

烷(3mL)溶液中，添加二苯基磷醯基疊氮化物(44.3mg，0.161mmol)與三乙胺(67.8mg，0.67mmol)。於室溫攪拌反應混合物30分鐘，然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(39.3mg，0.2mmol)。加熱反應混合物至100℃ 2小時。LC-MS顯示耗盡起始原料並檢測到所需質量。將其濃縮，利用矽膠管柱層析法，以於石油醚中之0%至100%乙酸乙酯洗提予以純化，得到1-[2-氯-8-[1-(二甲胺基)乙基]咪唑并[1,2-b]嗒

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(5.7mg，9.22%產率)，為白色固體。 To 2-chloro-8-[1-(dimethylamino)ethyl]imidazo[1,2- b ]ta

-7-carboxylic acid 9r (36mg, 0.134mmol) of 1,4-bis

To the alkane (3 mL) solution, diphenylphosphoryl azide (44.3 mg, 0.161 mmol) and triethylamine (67.8 mg, 0.67 mmol) were added. The reaction mixture was stirred at room temperature for 30 minutes, and then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (39.3 mg, 0.2 mmol) was added. The reaction mixture was heated to 100°C for 2 hours. LC-MS showed that the starting material was consumed and the required mass was detected. It was concentrated and purified by silica gel column chromatography with 0% to 100% ethyl acetate in petroleum ether to obtain 1-[2-chloro-8-[1-(dimethylamino) Ethyl]imidazo[1,2- b ]ta

-7-yl]-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (5.7 mg, 9.22% yield) as a white solid.

LC-MS: m/z 461[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.59(s, 2H), 9.24(s, 1H), 8.60(d, J = 2.0Hz ,1H),8.50(d, J =2.4Hz,1H),8.35(s,1H),8.15(s,2H),4.10-4.05(m,1H),2.30(s,6H),1.39(d, J = 6.4Hz, 3H).

-8-基]乙醯胺之合成 Example 68. N -[7-[[5-Chloro-6-(triazol-2-yl)-3-pyridyl]carboxamido]imidazo[1,2- b ]

Synthesis of -8-yl]acetamide

Example 68 was prepared in a procedure similar to that described in Preparation Example 3 by using 2-(trifluoromethyl)pyridine-4-amine as the starting material.

LC-MS: m/z 385[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 9.99 (brs, 1H), 8.95 (brs, 1H), 8.86 (s, 1H), 8.56 ( d, J =5.2Hz,1H),8.38(s,1H),8.07(d, J =1.6Hz,1H),7.62(dd, J =2.0,5.6Hz,1H),2.93(q, J =7.6 Hz, 2H), 1.24 (t, J = 7.6 Hz, 3H).

-7-基]脲/(S)-1-(5-氯-6-甲氧基-3-吡啶基)-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲/(R)-1-(5-氯-6-甲氧基-3-吡啶基)-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲之合成 Examples 69 to 71. 1-(5-Chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[1,2-b]

-7-yl]urea/( S )-1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[1,2 -b] Da

-7-yl]urea/( R )-1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[1,2 -b] Da

Synthesis of -7-yl]urea

Examples 69 to 71 were prepared similarly to the procedures described in the preparation of Examples 61 to 63, by using 5-chloro-6-methoxy-pyridin-3-amine as the starting material.

-7-基]脲(69)(30mg，21.04%產率)，為白色固體。 The title compound 1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[1,2-b]

-7-yl]urea ( 69 ) (30 mg, 21.04% yield) as a white solid.

LC-MS: m/z 377[M+H] ⁺ ; ¹ H NMR (400MHz; DMSO- d6 ): δ 9.95(s, 1H), 9.12(s, 1H), 8.59(s, 1H), 8.17( d, J =1.6Hz,2H),8.13(d, J =2.4Hz,1H),7.63(d, J =0.8Hz,1H),5.33-5.30(m,1H),3.92(s,3H), 3.31 (s, 3H), 1.53 (d, J = 6.4 Hz, 3H).

Using Prep-Chiral-SFC with the following conditions (column: Eclipse XDB-C18, column size: 4.6*150mm*5um, mobile phase: B (CAN), A (0.1% TFA)), chiral separation of compounds ( 69 ). The parts are evaporated to dryness to obtain the desired compound.

-7-基]脲(70)或(R)-1-(5-氯-6-甲氧基-3-吡啶基)-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲(71)。 Peak 1: ( S )-1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl]urea ( 70 ) or ( R )-1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[ 1,2-b]

-7-yl]urea ( 71 ).

Residence time: 9.290 minutes, ee >99%.

-7-基]脲(71)或(S)-1-(5-氯-6-甲氧基-3-吡啶基)-3-[8-(1-甲氧基乙基)咪唑并[1,2-b]嗒

-7-基]脲(70)。 Peak 2: ( R )-1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[1,2-b]ta

-7-yl]urea ( 71 ) or ( S )-1-(5-chloro-6-methoxy-3-pyridyl)-3-[8-(1-methoxyethyl)imidazo[ 1,2-b]

-7-yl]urea ( 70 ).

Retention time: 9.305 minutes, ee >99%.

LC-MS: m/z 377[M+H] ⁺ ; ¹ H NMR (400MHz; DMSO- d6 ): δ 9.95(s, 1H), 9.11(s, 1H), 8.59(s, 1H), 8.16( d, J =1.6Hz,2H),8.13(d, J =2.4Hz,1H),7.63(d, J =0.8Hz,1H),5.33-5.30(m,1H),3.92(s,3H), 3.31 (s, 3H), 1.53 (d, J = 6.4 Hz, 3H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 72. 1-(8-(1-Methoxyethyl)imidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

By using 6-(trifluoromethyl)pyridin-3-amine as the starting material, Example 72 was prepared in a similar procedure to that described in Preparation Example 61.

LC-MS: m/z 381[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 10.63(s, 1H), 9.08(s, 1H), 8.77(s, 1H), 8.58( d, J =5.6Hz,1H),8.20(d, J =1.2Hz,1H),8.07(d, J =1.6Hz,1H),7.67(d, J =1.2Hz,1H),7.63(dd, J = 2.0, 5.6 Hz, 1H), 5.31 (q, J = 6.8 Hz, 1H), 3.32 (s, 3H), 1.54 (d, J = 6.8 Hz, 3H).

-7-基)脲之合成 Example 73. 1-(2-(Difluoromethyl)pyridin-4-yl)-3-(8-(1-methoxyethyl)imidazo[1,2-b]

-7-yl) urea synthesis

Example 73 was prepared in a similar procedure to that described in Preparation Example 61 by using 6-(difluoromethyl)pyridin-3-amine as the starting material.

LC-MS: m/z 363[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 10.49(s, 1H), 9.08(s, 1H), 8.73(s, 1H), 8.50( d, J = 6.0Hz, 1H), 8.20 (d, J = 1.2 Hz, 1H), 7.89 (d, J = 1.6 Hz, 1H), 7.66 (d, J = 1.2 Hz, 1H), 7.54 (d, J = 5.2 Hz, 1H), 5.31 (q, J = 6.8 Hz, 1H), 3.32 (s, 3H), 1.55 (d, J = 6.8 Hz, 3H).

-7-基)脲/(S)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(8-(1-乙氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲/(R)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(8-(1-乙氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲之合成 Examples 74 to 76.1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(1-ethoxyethyl) Base) imidazo[1,2-b]ta

-7-yl)urea/( S )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(1 -Ethoxyethyl)imidazo[1,2-b]ta

-7-yl)urea/( R )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(1 -Ethoxyethyl)imidazo[1,2-b]ta

-7-yl) urea synthesis

By using different starting materials, Examples 74 to 76 were prepared in a similar manner as described in the preparation of Examples 61 to 63.

LC-MS: m/z 428[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d6 ): δ 10.58(s, 1H), 9.11(s, 1H), 8.76(s, 1H), 8.57( d, J =2.4Hz,1H), 8.50(d, J =2.0Hz,1H), 8.20(d, J =0.8Hz,1H), 8.16(s,2H), 7.66(d, J =1.2Hz, 1H), 5.44 (q, J = 6.4 Hz, 1H), 3.53 (m, 2H), 1.56 (d, J = 6.4 Hz, 3H), 1.21 (q, J = 8.8 Hz, 3H).

Using Prep-Chiral-SFC with the following conditions (column: Daicel Chiralpak, IG 4.6mm*250mm*5um, 5 to 50% Hex in EtOH, 1.0 mL/min), compounds were chirally separated ( 74 ). The parts are evaporated to dryness to obtain the desired compound.

-7-基)脲(75)或(R)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(8-(1-乙氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲(76)。 Peak 1: ( S )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(1-ethoxy) Ethyl) imidazo[1,2-b]

-7-yl)urea ( 75 ) or ( R )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8 -(1-Ethoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 76 ).

Residence time: 6.18 minutes; ee >99%.

-7-基)脲(76)或(S)-1-(5-氯-6-(2H-1,2,3-三唑-2-基)吡啶-3-基)-3-(8-(1-乙氧基乙基)咪唑并[1,2-b]嗒

-7-基)脲(75)。 Peak 2: ( R )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(1-ethoxy) Ethyl) imidazo[1,2-b]

-7-yl)urea ( 76 ) or ( S )-1-(5-chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8 -(1-Ethoxyethyl)imidazo[1,2-b]ta

-7-yl)urea ( 75 ).

Residence time: 7.70 minutes; ee >99%.

-7-基)脲之合成 Example 77. ( S )-1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(8-(1-methoxy Ethyl) imidazo[1,2-b]

-7-yl) urea synthesis

-7-基]脲(100mg，241.65μmol，得自實施例61至63之峰1)、 碳酸銀(2.0mg，12.08μmol)、與四氟硼酸鹽氟試劑(selectfluor)(34.2mg，96.66μmol)之乙腈(10mL)混合物。將反應管密封，微波輻射下，於85℃加熱，攪拌1小時。通過矽藻土墊以CH₂Cl₂過濾反應混合物，真空濃縮合併之有機層。所得混合物利用矽膠層析法(以於CH₂Cl₂中0至20%之MeOH洗提)，得到粗產物，然後以製備型HPLC(移動相：ACN-H₂O(0.05% HCO₂H)；梯度：40)純化，得到1-[5-氯-6-(三唑-2-基)-3-吡啶基]-3-[3-氟-8-[(1S)-1-甲氧基乙基]咪唑并[1,2-b]嗒

-7-基]脲(12.5mg，28.95μmol，11.98%產率)。 In a microwave tube, stir 1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-[(1S)-1-methoxyethyl]imidazo [1,2-b]

-7-yl]urea (100 mg, 241.65 μ mol, peak 1 from Examples 61 to 63), silver carbonate (2.0 mg, 12.08 μ mol), and tetrafluoroborate fluorine reagent (selectfluor) (34.2 mg, 96.66 μ mol) of acetonitrile (10 mL) mixture. The reaction tube was sealed, heated at 85°C under microwave irradiation, and stirred for 1 hour. The reaction mixture was filtered through a pad of _{celite with CH 2} Cl ₂ and the combined organic layer was concentrated in vacuo. The resulting mixture was subjected to silica gel chromatography (eluted _{with 0 to 20% MeOH in CH 2} Cl ₂ ) to obtain the crude product, which was then subjected to preparative HPLC (mobile phase: ACN-H ₂ O (0.05% HCO ₂ H) ; Gradient: 40) Purification to obtain 1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[3-fluoro-8-[(1S)-1-methoxy Ethyl]imidazo[1,2-b]ta

7-yl] urea (12.5mg, 28.95 μ mol, 11.98 % yield).

LC-MS: m/z 431.8[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ) δ 10.64(s, 1H), 9.18(s, 1H), 8.84(s, 1H), 8.56( d, J =2.3Hz,1H), 8.51(d, J =2.3Hz,1H), 8.17(s,2H), 7.50(d, J =7.1Hz,1H), 5.28(q, J =6.7Hz, 1H), 3.33 (s, 3H), 1.56 (d, J = 6.7 Hz, 3H).

-7-基)脲之合成 Example 78. ( R )-1-(5-Chloro-6-(2H-1,2,3-triazol-2-yl)pyridin-3-yl)-3-(3-fluoro-8-( 1-methoxyethyl) imidazo[1,2-b]

-7-yl) urea synthesis

-7-基]脲(得自實施例61至63之峰2)作為起始物質，以類似製備實施例77所述程序製備實施例78。 By using 1-[5-chloro-6-(triazol-2-yl)-3-pyridyl]-3-[8-[(1R)-1-methoxyethyl]imidazo[1, 2-b]

-7-yl]urea (from peak 2 of Examples 61 to 63) was used as the starting material, and Example 78 was prepared by a procedure similar to that described in Preparation Example 77.

LC-MS: m/z 431.8[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ) δ 10.63(s, 1H), 9.19(s, 1H), 8.82(s, 1H), 8.56( d, J =2.3Hz,1H),8.51(d, J =2.3Hz,1H),8.17(s,2H), 7.52(d, J =7.0Hz,1H),5.28(q, J =6.7Hz, 1H), 3.34 (s, 3H), 1.56 (d, J = 6.7 Hz, 3H).

-7-基)-3-(2-(三氟甲基)吡啶-4-基)脲之合成 Example 79. 1-(2-Chloro-8-(1-methoxyethyl)imidazo[1,2-b]

Synthesis of -7-yl)-3-(2-(trifluoromethyl)pyridin-4-yl)urea

Example 78 was prepared in a similar procedure to that described in Preparation Example 61 by using 2-(trifluoromethyl)pyridine-4-amine as the starting material.

LC-MS: m/z 415[M+H] ⁺ ; ¹ H NMR (400MHz; DMSO- d6 ): δ 10.68(s, 1H), 9.17(s, 1H), 8.83(s, 1H), 8.58( d, J =5.6Hz,1H),8.40(s,1H),8.06(s,1H),7.63-7.61(m,1H),5.22-5.17(m,1H),3.32(s,3H),1.53 (d, J = 6.8Hz, 3H).

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲之合成 Example 80. 1-[2-Chloro-8-[Methyl(methylsulfonyl)amino]imidazo[1,2- b ]

Synthesis of -7-yl]-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea

step 1

-7-羧酸乙酯7c(300mg，985.13μmol)、甲磺醯胺(140.6mg，1.48mmol)與碳酸二銫(641.9mg，1.97mmol)之1,4-二

烷(10 mL)混合物脫氣並再灌滿N₂(三次)。於90℃加熱該混合物並攪拌12小時。TLC顯示反應完全。以EA(50mL)稀釋該混合物，以飽和NH₄Cl(15mL x 2)洗滌並濃縮。殘留物利用快速矽膠層析法(以己烷/乙酸乙酯=1/1洗提)純化，得到2-氯-8-(甲磺醯胺基)咪唑并[1,2-b]嗒

-7-羧酸乙酯(230mg，產率：73%)，為白色固體。 Make 8-bromo-2-chloro-imidazo[1,2-b]

-7-carboxylic acid ethyl ester 7c (300mg, 985.13 μ mol), tosylamide (140.6mg, 1.48mmol) and dicesium carbonate (641.9mg, 1.97mmol) of 1,4-bis

The alkane (10 mL) mixture was degassed and refilled with N ₂ (three times). The mixture was heated at 90°C and stirred for 12 hours. TLC showed that the reaction was complete. The mixture was diluted with EA (50 mL), _{washed with saturated NH 4} Cl (15 mL x 2) and concentrated. The residue was purified by flash silica gel chromatography (eluted with hexane/ethyl acetate=1/1) to obtain 2-chloro-8-(methylsulfonamido)imidazo[1,2-b]ta

Ethyl -7-carboxylate (230 mg, yield: 73%), a white solid.

LC-MS: m/z 319.0 [M+H] ⁺ .

Step 2

-7-羧酸乙酯8s(120mg，0.376mmol)之N,N-二甲基甲醯胺(5mL)溶液中，添加碳酸鉀(104mg，0.753mmol)與碘甲烷(267mg，1.88mmol)，於80℃攪拌所得混合物2小時。LC-MS顯示反應完全。以乙酸乙酯(20mL)稀釋該混合物，以1N LiCl洗滌並濃縮。殘留物利用快速矽膠管柱層析法(以己烷類/乙酸乙酯=2/1洗提)純化，得到2-氯-8-[甲基(甲磺醯基)胺基]咪唑并[1,2-b]嗒

-7-羧酸乙酯(120mg，產率：96%)，為白色固體。 At room temperature, in 2-chloro-8-(methylsulfonamido)imidazo[1,2-b]ta

To the N,N-dimethylformamide (5mL) solution of ethyl-7-carboxylate 8s (120mg, 0.376mmol), potassium carbonate (104mg, 0.753mmol) and methyl iodide (267mg, 1.88mmol) were added, The resulting mixture was stirred at 80°C for 2 hours. LC-MS showed that the reaction was complete. The mixture was diluted with ethyl acetate (20 mL), washed with 1N LiCl and concentrated. The residue was purified by rapid silica gel column chromatography (eluted with hexanes/ethyl acetate=2/1) to obtain 2-chloro-8-[methyl(methylsulfonyl)amino]imidazo[ 1,2-b]

Ethyl -7-carboxylate (120 mg, yield: 96%), a white solid.

LC-MS: m/z 333.0 [M+H] ⁺ .

Step 3

-7-羧酸乙酯8t(120mg，0.36mmol)之THF(3mL)與水(1mL)溶液中，添加氫氧化鋰(34.5mg，1.44mmol)，於室溫攪拌反應12小時。TLC顯示反應完全。減壓去除溶劑，使殘留物溶於水(10mL)中。The溶液以1N HCl酸化該溶液至pH=6.0。過濾形成之沉澱，乾燥，得到2-氯-8-[甲基(甲磺醯基)胺基]咪唑并[1,2-b]嗒

-7-羧酸(85mg，77%產率)，為黃色固體，直接用於下一步驟。 To 2-chloro-8-[methyl(methylsulfonyl)amino]imidazo[1,2-b]ta

To a solution of 8t (120 mg, 0.36 mmol) of ethyl -7-carboxylate in THF (3 mL) and water (1 mL), lithium hydroxide (34.5 mg, 1.44 mmol) was added, and the reaction was stirred at room temperature for 12 hours. TLC showed that the reaction was complete. The solvent was removed under reduced pressure, and the residue was dissolved in water (10 mL). The solution was acidified with 1N HCl to pH=6.0. The precipitate formed is filtered and dried to obtain 2-chloro-8-[methyl(methylsulfonyl)amino]imidazo[1,2- b ]

-7-carboxylic acid (85 mg, 77% yield), as a yellow solid, was used directly in the next step.

LC-MS: m/z 305.1 [M+H] ⁺ .

Step 4

-7-羧酸9s(85mg，0.279mmol)之1,4-二

烷(5mL)溶液中，添加[疊氮(苯氧基)磷醯基]氧基苯(99.8mg，0.363mmol)與N,N-二乙基乙胺(141.1mg，1.39mmol)。所得混合物於此溫度攪拌30分鐘。然後添加5-氯-6-(三唑-2-基)吡啶-3-胺(81.9mg，0.418mmol)並於100℃攪拌混合物2小時。LC-MS顯示反應完全，以EA(20mL)猝滅，以飽和NH₄Cl(10mL x 2)洗滌。濃縮有機層，殘留物利用製備型TLC(DCM：MeOH=20：1)純化，得到粗產，，利用製備型HPLC將其純化，得到1-[2-氯-8-[甲基(甲磺醯基)胺基]咪唑并[1,2-b]嗒

-7-基]-3-[5-氯-6-(三唑-2-基)-3-吡啶基]脲(8.9mg，6%產率)，為白色固體。 At room temperature, in 2-chloro-8-[methyl(methylsulfonyl)amino]imidazo[1,2- b ]

-7-carboxylic acid 9s (85mg, 0.279mmol) of 1,4-bis

To the alkane (5 mL) solution, [Azide (phenoxy) phosphatidyl] oxybenzene (99.8 mg, 0.363 mmol) and N , N -diethylethylamine (141.1 mg, 1.39 mmol) were added. The resulting mixture was stirred at this temperature for 30 minutes. Then 5-chloro-6-(triazol-2-yl)pyridin-3-amine (81.9 mg, 0.418 mmol) was added and the mixture was stirred at 100°C for 2 hours. LC-MS showed that the reaction was complete, it was quenched with EA (20 mL) and _{washed with saturated NH 4} Cl (10 mL x 2). The organic layer was concentrated, and the residue was purified by preparative TLC (DCM:MeOH=20:1) to obtain a crude product, which was purified by preparative HPLC to obtain 1-[2-chloro-8-[methyl(methanesulfonate) (Amino)amino]imidazo[1,2- b ]ta

-7-yl]-3-[5-chloro-6-(triazol-2-yl)-3-pyridyl]urea (8.9 mg, 6% yield) as a white solid.

LC-MS: m/z 497.0[M+H] ⁺ ; ¹ H NMR (400MHz, DMSO- d ₆ ): δ 10.50(s, 1H), 9.43(s, 1H), 9.01(s, 1H), 8.55 (d, J =2.4Hz,1H),8.50(d, J =2.0Hz,1H),8.48(s,1H),8.17(s,2H),3.40(s,3H),3.33(s,3H) .

Claims (24)

A compound represented by structural formula (I) or a pharmaceutically acceptable salt or stereoisomer thereof,

among them A ring is

or

； Z is O, NR ₆ , or S; A ₁ and A ₂ are each independently CR ₁ or N; Each case of R ₁ is hydrogen; halogen; -OH; CN; -COOC _1-6 alkyl; if necessary, C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; phenyl; amine group; N, N - two -C _1-6 alkylamino; is optionally substituted by halogen, phenyl, or substituted heteroatoms selected from N and O of the 1-3 5-6 heterocycle of C _{1 -6} alkyl, the ring of the heterocyclic ring is _{optionally substituted by C 1-6} alkyl; Rh; ORh; a 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, which ring is optionally required _{C 1-6} alkyl substituted by amino, optionally amino or hydroxy, or N -mono-C _1-6 alkylaminocarbonyl or N,N -di-C _1-6 alkylamino Carbonyl substitution; where Rh is a 5- to 6-membered heterocyclic ring having 1 to 4 heteroatoms selected from N, O and S, and the ring _{is optionally substituted with C 1-6} alkyl, -OH, or pendant oxy, Each case of R ₂ is hydrogen; halogen; CN; -COOC _1-6 alkyl; optionally C _1-6 alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; amino; N,N- two -C _1-6 alkylamino; is optionally substituted by halogen, -OH, a phenyl group, or selected from N and O substituted with 1-2 of the 5-6 atom of the heterocycle C _1-6 alkyl The ring of the heterocyclic ring is _{optionally substituted with a C 1-6} alkyl group; Rh; ORh; a 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, and the ring is optionally substituted by an amino group , If necessary, C _1-6 alkyl substituted by amino or hydroxy, or substituted by N -mono-C _1-6 alkylaminocarbonyl or N,N -di-C _1-6 alkylaminocarbonyl; Each case of R ₃ is H; deuterium; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; -C(=O)NR _a R _b ; optionally halogen, -OH, C _1-6 alkyl, -NH ₂ , -NHC(=0)C _1-6 alkyl, N -di-C _1-6 alkylamino, or N -C _1-6 alkoxy substituted by mono-C _1-6 alkylamino group; optionally halogen, C _2-6 alkenyl, -OH, -NH ₂ , -NHC(=O)C _1-6 Alkyl, N -di-C _1-6 alkylamino, N -mono-C _1-6 alkylamino, -O-Rg, Rg, phenyl, or C _1-6 alkoxy substituted C _1-6 alkyl, where the alkoxy group is optionally substituted by halogen, -OH, C _1-6 alkoxy, N,N -di-C _1-6 alkylamino, Rg or phenyl; C _3-6 cycloalkane substituted by halogen, -OH, C _1-6 alkyl, N,N -di-C _1-6 alkylamino or C _1-6 alkoxy-C _{1-6 alkyl} Group; phenyl substituted with halo or C _1-6 alkoxy as required; 5- to 6-membered heteroaryl ring with 1 to 3 heteroatoms selected from N and O, and the ring may be optionally controlled Amino or -OH-substituted C _1-6 alkyl substitution; Rg; or N,N -di-C _1-6 alkylaminocarbonyl; wherein Rg is a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, and the ring is optionally controlled by -OH, -NH ₂ , C _1-6 alkyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution; R _a is independently H or an optionally substituted C _1-6 alkoxy group of C _1-6 alkyl, R _b is independently H, C _1-6 alkyl, -COC _1-6 alkyl, -SO ₂ C _1-6 alkyl, C _3-6 cycloalkyl or a 3- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N and O, the ring is optionally connected to C _1-6 alkyl, C _{1 -6} alkoxy-C _1-6 alkyl, or C _1-6 alkoxy-carbonyl substitution, or R _a and R _b together with the nitrogen atom to which it is connected, form a 4- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O, and S. The ring is optionally controlled by -OH, -NH ₂ , N -Di-C _1-6 alkylamino, N -mono-C _1-6 alkylamino, C _1-6 alkyl, C _1-6 haloalkyl, C _1-6 alkoxy, C _{1 -6} haloalkoxy, O-cyclopropyl, C _1-6 alkoxy-C _1-6 alkyl, or C _1-6 alkyl-carbonyl substitution; Each case of R _{c is C 1-6} alkyl or C _3-6 cycloalkyl; Wherein the R _a, R _b, or R _c or as shown by the R _a, R _b, or the R groups depending on the alkyl group shown _c needs halogen, -OH, C _1-2 alkoxy, or C _3-4 cycloalkyl substitution; Each case of R ₄ is H, deuterium, halogen, CN, C _1-6 alkyl, or C _1-6 haloalkyl; Each case of R ₄ 'is H, deuterium, F, or Cl; Each case of R ₅ is H, deuterium, C _1-6 alkyl, or C _1-6 haloalkyl; and R ₆ is H; OH; optionally substituted with halogen, OH, C _1-6 alkoxy or C _1-6 alkyl; or an optionally halogen-substituted OH, C _1-6 alkoxy groups or C _3-6 cycloalkyl. The compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Z is O. The compound according to claim 1 or 2, or a pharmaceutically acceptable salt or stereoisomer thereof, wherein the A ring is

. The compound according to any one of claims 1 to 3, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein _{each case of R 3} is H; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; -C(=O)NR _a R _b ; optionally halogen, -OH, C _1-6 alkyl, -NH _2. -NHC(=O)C _1-6 alkyl, N -di-C _1-6 alkylamino, or N -mono-C _1-6 alkylamino substituted C _1-4 alkoxy ; is optionally substituted by halogen, C _1-2 alkoxy, N - two -C _1-6 alkylamino, or N - the mono-substituted -C _1-6 alkylamino C _1-4 alkyl; C _3-6 cycloalkyl; or Rg. The compound according to any one of claims 1 to 4, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein the compound is represented by structural formula (II-A) or (II-B) :

Or a pharmaceutically acceptable salt or stereoisomer thereof, wherein _{each case of R 4} ′ is H or F. The compound according to any one of claims 1 to 5, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein Examples of R ₃ are H; halogen; CN; -OH; -COOH; -NR _a R _b ; -SR _c ; -SO ₂ R _c ; -SO ₂ NR _c ; -C(=O)NR _a R _b ; is optionally substituted by halogen, -OH, C _1-4 alkyl, -NH _2, N - two -C _1-4 alkylamino, or N - mono-substituted -C _1-4 alkyl group of C _{1 -4} alkoxy; is optionally substituted by halogen, C _1-2 alkoxy, N - two -C _1-4 alkylamino, or N - mono-substituted -C _1-4 alkyl group of C _{1- 4} alkyl; C _3-6 cycloalkyl; morpholino; oxetanyl; or azetidinyl substituted by methyl as required; R _a is independently H or an optionally substituted C _1-4 alkoxy group, the C _1-4 alkyl; R _{b is} independently H, C _1-4 alkyl, -COC _1-4 alkyl, -SO ₂ C _1-4 alkyl, C _3-6 cycloalkyl or having 1 to 3 selected from N and O A 3- to 6-membered heterocyclic ring with three heteroatoms, which is optionally substituted _{by C 1-4} alkyl, C _{1-4 alkoxy-C 1-4} alkyl, or C _1-4 alkoxy-carbonyl, or R _a and R _b , together with the nitrogen atom to which they are connected, form a 4- to 6-membered heterocyclic ring having 1 to 3 heteroatoms selected from N, O, and S, and the ring is optionally controlled by -OH, C _1-4 Alkyl, C _1-4 haloalkyl, C _1-4 alkoxy, C _1-4 haloalkoxy, or C _1-4 alkoxy-C _1-4 alkyl substituted. The compound according to any one of claims 1 to 6, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein _{each case of R 4} is H, halogen, and C substituted by a halogen group as necessary _1-2 alkyl, and each case of _{R 4 ′ is H.} The compound according to any one of claims 1 to 7, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₂ is H, Cl, CN, or optionally substituted by halogen or -OH的C _1-6 alkyl. The compound according to any one of claims 1 to 8, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein _{each case of R 1} is H; halogen; -OH; CN; _{C 1-6} alkoxy substituted by halogen; C _1-6 alkoxycarbonyl; phenyl; N,N -di-C _{1-6 alkylamino; C 1-} substituted by halogen or phenyl if necessary ₆ alkyl group; a 5- to 6-membered heteroaryl ring containing 1 to 3 N atoms, the ring may optionally be optionally substituted with an amino group or a hydroxy group C _1-6 alkyl group or mono- N -C _1-6 Alkylaminocarbonyl or di- N -C _1-6 alkylaminocarbonyl substituted; ORh; or Rh; wherein Rh is a 5- to 6-membered heterocycle containing 1 to 4 heteroatoms selected from N, O and S Cyclic group, the ring _{is optionally substituted with C 1-6} alkyl, -OH, or pendant oxy. The compound according to any one of claims 1 to 9, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₁ is H; halogen; C _1-4 alkane substituted with halogen as necessary Group; C _1-4 alkoxy; or 5- to 6-membered heteroaryl containing 1 to 3 N atoms. The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt, or a stereoisomer thereof, wherein _{each case of R 3} is bromo, methyl, ethyl, propyl, iso Propyl, COOH, -CH(CH ₃ )OCH ₃ , -CH(CH ₃ )OCH ₂ CH ₃ ,

,

,

,

, -CH(CH ₃ )N(CH ₃ ) ₂ , -CH ₂ OCH ₃ , cyclopropyl, morpholinyl, -S(CH ₃ ), -N(CH ₃ ) ₂ , -N(CH ₃ )( CH ₂ CH ₃ ), -N (CH ₂ CH ₃ ) ₂ , -N (CH ₃ ) (isopropyl), -N (CH ₃ ) (cyclopropyl), -N (CH ₂ CH ₃ ) (ring Propyl), -N(CH ₂ CH ₂ OCH ₃ ) (cyclopropyl), -N(CH ₃ ) (SO ₂ CH ₃ ), or -N(CH ₃ ) (CH ₂ CH ₂ OCH ₃ ). The compound according to any one of claims 1 to 10, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein _{each example of R 3} is -CH(CH ₃ )OCH ₃ ,

,

, Ethyl, or cyclopropyl. The compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₄ is H, F, Cl, Br, CH ₃ , or CF ₃ . The compound according to any one of claims 1 to 13, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₂ is Cl or CF ₃ . The compound according to any one of claims 1 to 14, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₁ is H, 1,2,3,-triazole, -OCH ₃ , Or CF ₃ . The compound according to any one of claims 1 to 15, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein R ₄ is H or Cl. The compound according to any one of claims 1 to 16, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, wherein

for

. A pharmaceutical composition comprising an effective amount of the compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt thereof, or a stereoisomer thereof, and a pharmaceutically acceptable carrier. A combination comprising a therapeutically effective amount of the compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt or stereoisomer thereof, and one or more co-agents with therapeutic activity. A method for inhibiting MALT1 activity in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the compound according to any one of claims 1 to 17, or pharmaceutically acceptable The salt or its stereoisomer; or the pharmaceutical composition according to claim 18; or the combination according to claim 19. The method of claim 20, wherein the subject has a disease or symptom such as an autoimmune disease, inflammatory disease, or cancer, and wherein the disease or symptom is treated for inhibiting MALT1 activity. The method according to claim 21, wherein the disease or symptom is rheumatoid arthritis (RA), multiple sclerosis (MS), systemic lupus erythematosus (SLE), vasculitis, allergic disease, Respiratory diseases [such as asthma and chronic obstructive pulmonary disease (COPD)], symptoms caused by delayed or immediate allergic reactions, acute allergies, acute or chronic transplant rejection, graft-versus-host disease, cancer of hematopoietic origin, or solid tumors [Including chronic myelogenous leukemia (CML), myelogenous leukemia, non-Hodgkin's lymphoma (NHL), or B-cell lymphoma]. The method according to claim 21 or 22, wherein compared with Treg, the disease or symptom is treated by selective or preferential inhibition of Th17. The compound according to any one of claims 1 to 17, or a pharmaceutically acceptable salt or stereoisomer thereof, is used as a medicament, for example, as a medicament for an MALT1 inhibitor. 0

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